Genetics

Topic 1 –DNA, Cell features, Cell cycle

 Diploid – 23 chromosomes with 46 chromatids , like somatic/autosomes
cell(meiosis1/mitotic)
 Haploid – 23 chromosomes with 23 chromatids, like gametes (meiosis 2)
 DNA bases - purines (two rings) – adenine and guanine| pyrimidines (one ring)
–Thymine and Cytosine.
Guanine goes with Cytosine (same percent of the strand), Adenine goes
Thymine (same percent of the strand).
In RNA – Thymine replaced Uracil. It's one Stranded and the sugar is ribose
 DNA strands are antiparallel, the bonds between sugars on adjacent nucleotides
are called phosphodiester.
 Essential attributes -
-Carry all of the info. Needed to direct the specific organization and metabolic
activates of the cell.
-Replicate accurately and have repair mechanism
-Capable of undergoing occasional mutations
 Chromosomes that pair in meiosis and same genetic loci and structure are
Homologues Chromosomes
 Paring of homologs chromosome in meiosis known as synapsis
 Nucleosome  Solenoid  Protein scaffold
 Chromatin – nucleosome its composed of Histone octamer about 140bp DNA
 Kinetochore - A structure at the centromere to which the spindle fibers are
attached. And takes part in the segregation of chromosome towards the poles.
 Nucleosome - The primary structural unit of chromatin, consisting of 146 base
pairs of DNA wrapped twice around a core of eight histone molecules.
 Human Mitochondrial genome – come from maternal DNA, Small and
circular, 16kb lengths encodes for 37 genes.
 The human genome – is 3 Billion bp but less than 1.5% code for proteins
 Difference between mitosis to meiosis – they both line up on the equatorial
plate but mitosis is a dyad consist of two chromatids while meiosis 2
homologous chromosome with tetrads consist 4 chromatids.
 Protein with most important dispersed repeated sequences Alu (10%) and Line
family (20%)
 Main accomplishments of Mitosis insure that each of the two daughter cells
receives a complete set of genetics info.
 Main accomplishments of meiosis generates diversity by
1. Crossing over
2.independent assortment as well insuring same number of chromosomes from
generation to generation.
 Cell go into mitosis has twice as much as genetic material than cell just finished
mitosis.
  Phases of Cell cycle are:
G1, S in which replication occur, G2 and M
 1st division of meiosis (same as mitosis) except in meiosis the homologues
chromosome separate and in mitosis sister chromatids
 In meiosis 2 – 46 chromatids separate to two cells with 23 chromatids
 In mitosis – 92 chromatids separate to two cells with 46 chromatids
 Chiasmata is the point where two homologous chromosome are held together
Topic 3- gene structure and function
 Enhancers are sequence elements that can act at a distance from gene to
stimulate transcription. Interaction with specific proteins leads to increased
levels of transcription
 Amino acyltRNA synthetase attach the accurate amino acid onto the
corresponding tRNA
 Ribosome Complex reads 3 mRNA base pair at a time in order to determine the
a.a, a shift in number of nucleotides isn’t divisible by 3 in the reading frame will
cause result in subsequent codons to be read differently. That’s why excision of
introns need to be precise.
 Alternative splicing – many genes the primary transcript can follow multiple
pathways leading to multiple related but different mRNA in each we can
generate different protein products.
Many genes capable producing different proteins
Post transitional modifications
those are the reasons for amount of genes is not equal to amount of proteins.
 RNA polymerase
I- used exclusively in producing the transcript becomes processed into ribosomal
RNA
II-responsible for transcribing all protein coding genes
III- transcribes all tRNA genes and the 5S component of rRNA
 Promoter – site of RNA polymerase binding to DNA template
 snRNP – molecule which specialized in splicing RNA when combined with
certain proteins
 Poly A' tail – added to the 3' end of mRNA molecule
 Exon shuffling – model of protein evolution with the combination of different
exons
 Transcription in eukaryotes occurs in nucleus and Translation occurs in
cytoplasm(ribosomes)
 Housekeeping genes – expressed all the time in low levels in all cells. (also
called Constitutive gene expression)
 In prokaryote - Translation and transcription occurs at the same time, there
are multiple sites for translation
there is no Exons and Introns and no G cap and poly a tail.
 DNA methylation alters gene expression the most frequent DNA modifications
 Chromatin remodeling complex allows access of condensed genomic DNA to
regulatory transcription machinery proteins and thereby controls gene expression
  In eukaryotic – there is exons and introns, each gene has its own
transcriptional control. Translation and transcriptions occurs separately
 Cis regulatory -sequences it’s a region of DNA or RNA that regulates the
expression of genes located on the same molecule
 Trans regulatory – sequences which may modify the expression of distant
genes. More specifically Trans – regulatory elements are DNA sequences that
encode transcription factors
 Pre-mRNA -> mRNA – slicing out introns, G cap is added to 5' end and poly a
tail is added to 3' end.
 Charged tRNA is tRNA that the correct amino acid added to its done by the
enzyme aminoacyl tRNA synthetase
 Pseudo-autosomal region - Segment of the X and Y chromosome, located at
the most distal portion of their respective p and q arms, at which crossing over
occurs during male meiosis. Traits due to alleles at pseudo-autosomal loci will
appear to be inherited as autosomal traits despite the physical location of these
loci on the sex chromosomes.
Topic 5&6 – Cytogenetics

Syndrome karyotype Symptom extra

Turner syndrome X,45 Wide neck, low-set ears, Monosomy for X
short(height) ,chromosome
No barr body
Can occur by non-
disjunction
Down syndrome XX,+21 or ,47 Mental retardation, short Trisomy 21
47,XY,+21 ,stature
short wide neck
Klinefelter XXY,47 ,Infertile Can occur by non-
syndrome learning difficulties disjunction
Edwards XX,+18 or ,47 ,Kidney malformation Trisomy 18
syndrome 47,XY,+18 heart defects, growth
deficiency
Patau syndrome XX,+13 or ,47 Heart defects, kidney Trisomy 13
47,XY,+13 defects, deformed feet
Robertsonian XX,-14.t(14;21) ,46 Mental retardation, short Trisomy 21 – down
translocation or 46,XY,- stature, short wide neck syndrome
14.t(14;21)
Cri De Chat -XX,5p,46 Cause mental retardation Deletion of short arm
syndrome
Triple X XXX,47 ,Tallness barr body 2
syndrome ,menstrual irregularities
slight impact on
intelligence
 Autosomal recessive • Cystic fibrosis (mutant chloride channel)
inheritance (AR) • Phenylketonuria (PKU)
• Tay-Sachs disease (gangliosidosis)
• Alkaptonuria
• Sickle cell disease
• Albinism

Autosomal dominant • Huntington disease (late onset lethal disease)

inheritance (AD) • Aachondroplasia
• Familial hypercholesterolemia
• Brachydactylia
• Neurofibromatosis
• Alzheimer-disease

X-linked recessive • Duchenne’s muscular dystrophy

inheritance (XR) • Testicular feminization syndrome
• G6PD deficiency
• Fragile X syndrome (not quite recessive)
• Haemophilia
• Color-blindness

X-linked dominant • Myotonic Dystrophy

inheritance (XD)
Y-linked inheritance (Y)

 Gain-of-function mutations may alter the biochemical phenotype by enhancing

one or more of the normal functions of a protein. A gain in protein function may
be due to either an increase in its expression, or an increase in the ability to
perform its biological function. E. g.:
Hb Kempsey – locks hemoglobin in its high oxygen affinity state, herby reducing
oxygen delivery to tissue.
Achondroplasia, Alzheimer disease.
 Loss-of-function mutations of a gene may result from alteration of its coding,
regulatory, or other critical sequences due to the introduction of nucleotide
substitutions, deletions, insertions, or rearrangements.
E.g.: α- or β-thalassemia – α due to deletion of globin genes
, retinoblastoma, phenylketonuria
 Tolerance for sex chromosomes -
Y- encode for only few genes
X- there is inactivation regardless of how many X chromosomes there until only 1
is
active.
 Types of abnormalities in chromosome structure –
Deletions
Duplication
 Translocations
Rings
 Types of viable trisomy's exist -
8,9,13,18,21,22, XXX, XXY, XYY
 Nondisjunction can explain chromosomal mosaicism when occurs during
meiosis 1,2 and during mitotic division
 Isochromosome its mis-divison trough the centromere when there is exchange
involve one arm of chromosome ad its homologue in the region of the arm
adjacent to the centromere
 Uniparental disomy – when two chromosomes inherited from only one parent
Exp. Uniparental disomy of chromosome 15 can be Angelman syndrome
(inherited only from the father) or Parder-wili syndrome (inherited only from the
mother).
 SRY- is the sex determine region Y that is located on the Y chromosome
 Xist (X-inactive specific transcript) - is an RNA gene on the X chromosome
that acts as a major effector of the X inactivation process. The Xist RNA
transcript is expressed on the inactive chromosome and not on the active one.
 Balanced Carrier/ Unbalanced Carrier – balanced lead to healthy individual
while unbalanced lead to unhealthy
 Chimerism- formed by:
Dispermic chimeras – result from fusion of two zygotes into one individual.
Blood group chimera - exchange of hematoietic stem cells by DZ twins in utero.
Its inevitable result of transplantation.
The difference from mosaicism which is the splitting of single zygote to
tissue/individual with two cell lines. It's an individual derived from two
genetically different zygotes.
 Robertsonian translocation - can affect only acrocentric chromosome ,and the
one can participate are 13,14,15,21 and 22

Topic 7 –Transmission genetics

 Recessive epistasis- when one gene masks the expression of another gene,
recessive is when the recessive homozygosity masks the expression of another
gene whether it dominant or recessive. Exp. Gene that causes albinism will mask
the color of person's hair.
 Linkage group – is a group of genes that located close to teach other on the
same chromosome and tend to be transmitted together.
 Pleiotropy – single gene controlling of influencing multiple phenotypic traits.
Exp. PKU can cause mental retardation and reduced hair and skin pigmentation,
can be caused by any of large number of mutations in single genes that codes for
an enzyme which converts the amino acid phenylalanine to tyrosine.
 Alleles – is the alternative types of gene
 Homozygote – is having the allele of given gene on homologous chromosome
 Heterozygote – is having the dominant and recessive allele on homologous
chromosome
 Testcross – mating heterozygous with recessive homozygote
 Phenotype – is physical appearance of a character
 Genotype – is genetic constitution
 Locus – is the region of chromosome occupied by a gene
 Penetrance – is the proportion of population with certain genotype that will
exhibit the associated phenotype
 Expressivity – it’s the severity of expression of the phenotype among
individuals with the same disease causing genotype
 Phenocopy – phenomenon in which the environmental conditions mimic
phenotype produced by a gene, for exp. Vanessa genus butterflies can change
phenotype based on the local temp.
 Genetic heterogeneity – in contrast to pleiotropy where a single gene may cause
multiple phenotypic expressions, this is a phenomenon in which single
phenotype or disorder are caused by any one of the multiple number of alleles,
exp. Alzheimer disease.
 Centi-morgan – is the unit of genetic distance, which determined by measuring
the rate or recombination in genetic crosses.
 Rate of recombination – we take both of small numbers add them together and
dived by all products then divided it by 2 and we got the rate of
recombination.
If it's equal its means there is no linkage between genes and its independent
assortment
 Monohybrid –doing a cross with only one gene (type of allele) exp. Only eyes
(Aa/AA/aa)
 Dihybrid – doing a cross with two genes that encode for different things
exp. Eyes and wings (AABB/AABb/AaBB/aaBb/Aabb/aabb)
The phenotypic ratio for both heterozygotes is always 9:3:3:1
unless there is recessive epistasis and then will see 9:3:4 | Complementary gene
activity then will see 9:7
 O blood type – can be formed by recessive homozygote genes or by H gene
recessive inheritance
 H gene – responsible for formation of H precursor on membrane of the RBC, if
we have hh genotype the precursor won't be formed and the phenotype will be O
blood even if we have alleles for A/B antigens.
 Rh – during pregnancy when mother is Rh- (means doesn’t have the Rh antigen)
and the fetus is Rh+ it can elicit immune response towards the fetus. In the first
pregnancy the mother doesn’t have the Rh antibodies and therefore the immune
system won't elicit a response against the fetus.
During the second pregnancy of Rh- mother with Rh+ fetus the mother's body will

attack the fetus because the Rh+ recognized as antigens by the antibodies
 Linkage disequilibrium – it’s the nonrandom association of alleles at two or
more loci that may or not be on the same chromosome. Basically shoes the
frequency of some combinations of alleles or genetic markers in a population.
 Compound (compound heterozygote) - An individual, or a genotype, with two
different mutant alleles at the same locus. Not to be confused with
homozygote, in which the two mutant alleles are identical.

Topic 8 - basis of complex inheritance

  Discontinues traits – mendelian monogenic determination of phenotypic traits
these genes are called major genes. The gene effect or genotype can be deduced
from phenotype
 Continuous traits – non mendelian (polygenic) determination of phenotype.
These genes called minor genes. The gene effect can't be deduced from
phenotype
 Meristic traits – trait in which the phenotype is determined by counting.
Exp. Number of eggs laid by hen.
 Exp. Of human threshold traits -
spina bifida, congenital dislocation of the hip, cleft lip and/or palate, pyloric
stenosis.
 Exp. Of polygenic system -
hyperlipidemia, familial hypercholesterolemia, diabetes mellitus.
 QTL (quantitative trait locus) – locus segregating for alleles that have
different, measurable effects on the expression of quantitative trait.
 Risk genes – when different forms of gene are known to be associated with
different levels of risk for specific health problem.
 Threshold trait – trait that have only two or few phenotypic classes, but the
inherited by effect of multiple genes acting together with the environment.

Topic 9 – DNA polymorphisms, mutation & repair

 Genetic marker – a gene or DNA sequence with a known location on a

chromosome, can be used identify cells, individuals or species.
exp. Classical markers ABO blood groups, MHC
exp. Molecular markers SSR's, RFLP's
 Important molecular methods in detection of DNA polymorphisms -
southern blot
electrophoresis
 Most important DNA polymorphisms -
SNP,RFLP,STR
 SNP and RFLP relationship- both are polymorphism can be used as genetic
markers. SNP are polymorphisms which arise via changes of single nucleotide.
RFLP are differences in fragment length produced by the presence or absence of
the cleavage sites in DNA molecules. This produce different lengths DNA
fragments.
Both can be used in DNA typing screening human DNA for presence of
potentially deleterious genes for exp. Sickle cell anemia
 Short tandem repeat polymorphisms (STRP) – used in DNA typing which is
practically important to forensic investigations being used to prove someone
connection to a crime.
 Silent mutations - are DNA mutations that don’t result in a change to the amino
acid sequence of a protein (due to redundant character of codons) or that result in
amino acid change but don’t result in radically different properties of the
changed amino acid.
 Nonsense mutation – point mutation in a DNA sequence which result in a
premature stop codon usually results in nonfunctional protein.
  Missense mutation – point mutation in which a shingle nucleotide changed
resulting in a codon that codes for a different amino acid, may render the protein
as non-functional.
 DNA damages – it means an error in the DNA sequence, it can be caused by
-Depurination the bond between the sugar and the base is broken and the base is
replaced by hydroxyl (OH).
-Demethylation/deamination those damages can be repaired but even if the
damage is repaired, the repair mechanism may read the complementary strand
not correctly and cause a mutation and the error will remain in the DNA after
cell division is permanent and basically it’s a mutation.
 Dynamic mutation (can be called pathogenic repeat expansion) – is an unstable
heritable element where probability of expression of mutant phenotype is
function of the number of copies of the mutation. Exp. Huntington disease,
fragile X syndrome.
 Micro-satellite markers – microsatellite are repeating units of 2-9 base pairs.
Used for human identification looking at genetic relatedness and DNA typing.
 Mini-satellite markers – is a section of DNA that consists of a shorts series of
bases 10-60 base pairs that occur more than 1000 locations in human genome.
 Bombay blood group –it’s a group in which there is no precursor for the
formation of A/B antigens because they have genotype of hh for the H gene.
Which means the antigen isn’t formed. Even if they have alleles for A or B.
person with Bombay blood group have antibodies for A,B and for the H antigens
 Major histocompatibility complex (MHC) – The complex locus on
chromosome 6p they are highly polymorphic, polygenic and co-dominant
expressed. Enables to distinguish between components of the body and foreign
ones which helps immune system.
 Haplotype – is the several alleles reside on a piece of a chromosome very
closely together so they are inherited as one unit.
 Methods for HLA typing – cellular typing, gene sequence, phenotyping
 Haploinsufficiency - A cause of genetic disease in which the contribution from
a normal allele is insufficient to prevent disease because of a loss-of function
mutation at the other allele.

Topic 9 – population genetics *‫לתרגל‬

 Hardy Weinberg principle – p+q=1, p2+2pq+q2=1

 Ideal population – contains:
no mutations
mating is random
population size is indefinite
no natural selection
 Gene drift – process in which chance determines the direction of unpredictable
changes of allele frequencies only in small population
 Random mating – individuals do not preferentially choose mates with certain
genotypes
 Assorting mating - individuals do preferentially choose mates with certain
genotypes
 Inbreeding – marriage between relatives in a pedigree or in population genetics
Topic 10 – Gene mapping


Topic – Hemoglobinopathies *‫לעבור על זה לעומק‬

 Novel property mutation – change in amino acid sequences caused disease by

giving novel property on the protein without necessarily altering its normal
function. Exp. Sickle cell anemia
 Mutation associated with ectopic or Heterochorinc gene expression –
mutations that alter the regulatory regions of gene to cause inappropriate
expression. Exp. Cancer is most common which is frequent due abnormal
expression of gene.
 Hemoglobin – oxygen carrier, contains four subunits (2 alpha, 2 beta) each
subunit composed of polypeptide chain. With 8 helical regions.
 Genes cluster codes for globin chains – for alpha chains is found on the
chromosome 16 there you have a1 and a2 which are identical. For beta chains is
found on chromosome 11 and there is only one beta gene.
 Red blood cell formation -
in embryo the first site is the yolk sac
later in embryonic life the liver is the main producer
in the human adult it's found in the bone marrow
fetal hemoglobin is predominant in HbF and adult hemoglobin is HbA 2 and
HbA
 Hemoglobin diseases- are classified into 3 categories
1.varients that cause hemolytic anemia
2.mutants with altered oxygen transport
3.varients due t mutations in coding region that cause thalassemia.
 Beta globin mutations- not all causes disease or responsible for the hemoglobin
beta gene.
 Alpha – thalassemia – caused reduction of a-globin production so it's
significantly reduced at birth. And by that cause newborns severe damage
 Beta-thalassemia – results from reduction in abundance of b-globin. Which is
one of the major adult hemoglobin proteins in RBC so even if the child appear
healthy at birth.
Treatment is by increase the synthesis of delta and gamma globin (only 2% of
total adult hemoglobin) in relating transcriptional factors can be possible
treatment.

Topic 11 - Bacterial genetics

 Plasmids – are circular DNA molecules which replicate independently from the
chromosome and found only in bacteria, F+ contains F plasmid which allow
genes to be transferred to a recipient bacteria, can insert itself into bacterial
 chromosome by homologous recombination. F- lacks the F plasmid and only
such bacteria can function as recipient cell.
 Three major types of genetic transfer
Transformation – when DNA molecule is taken up from external environment and
incorporated into the genome.
Conjugation – when donor DNA is transferred from one bacterial cell to another
by plasmid during direct content.
Transduction – when DNA is transferred by a bacteriophage to another cell.
 Auxotroph – microorganism unable to synthesize an essential compound but
able to grow if it's supplied by exogenously. The mutant of auxotroph require
metabolite that isn’t needed by the wild type.
 Prototroph –wild type bacterial strain capable of growth in defined minimal
medium contain only carbon source and inorganic compounds.
 Lysogen – is a cell which carry the prophage which is the integrated phage DNA
 Lytic cycle – is a series of events following infection or induction by prophage
or temperate phage which include DNA replication  formation of mature
phage  and the lysis (killing) of cell.
 Utilization – differs from the wild type by their ability/inability to use specific
compounds such as carbon or nitrogen for growth.
 Coupled transcription- translation – means that translation begins while
mRNA is still synthesized.
 Attenuator – is provisional stop signal located in the DNA segment that
corresponds to leader sequence of mRNA
 E.coli – the operator is a short region of DNA that lays partially within the
promoter and interacts with regulatory protein that controls transcription of the
operon.
Lac operon in uneducable when glucose is present cause it will be energetically
wasteful to express lac genes.
when the cell is growing in the presence of lactose the cAMP levels are high and
cAMP-CRP is positive regulator of lac operon, the negative regulator is Lac I
trp is negative repressive regulation.
 Polycistorinc – An mRNA molecule which contain coding sequences for several
polypeptide chains.

Topic 12 – Molecular basis of genetic diseases

 Housekeeping proteins – present in virtually every cell and have fundamental

roles in maintenance of cell structure and function. More genes coding these
proteins in RNA (90%). More in housekeeping gene happens more frequently.
 Tissue specific specially proteins – produced in only one or limited number of
cell types and have unique functions that contribute to the individually of the
cells in which they expressed. Less genes coding these proteins in RNA (10%)
 Variable of genetic diseases- is due to one of those
1.allelic heterogeneity - presence of multiple alleles at a locus
2.locus heterogeneity –arise from the association of one or more than one locus
with a specific clinical condition, situation termed locus heterogeneity
3.effect of modifier genes-phenotypic variation can be due environmental factors
or the action of other genes are termed as modifier genes.
 Storage disease – due to genetic defect of a variety of biological enzymes
  Ecogenetics – the influence the environment has on gene expression
 Type of proteins affected in single-gene genetic disorders -
receptor protein, structural protein, transport proteins, neuronal proteins.
 Cystic fibrosis – CFTR gene undergoes mutation, deletion of phenylalanine
residue which results in loss of function of CFTR protein which is involved in
fluid and electrolyte transport across epithelial apical membranes. Its autosomal
recessive. Lungs and exocrine pancreas are affected.
 Duchenne muscular dystrophy – deletion in gene that codes for DMD gene,
its x-linked recessive so males are usually most affected. Dystrophin link the
ECM to actin. Without it muscle membrane integrity can't be maintained.
Less likely female will occur, but it can happen by nonrandom X-inactivation,
turner syndrome with mutation on X-chromosome, or X translocation, and rarely
heterozygous monozygotic twins.
By screening for deletions and duplications by multiplex polymerase chain
reaction we are able to diagnose DMD parentally.
 Anticipation -The progressively earlier onset and increased severity of certain
diseases in successive generations of a family. Anticipation is caused by
expansion of the number of unstable repeats within the gene responsible for the
disease.
 Dominant negative effect – means there is an altered gene product that impair
the wild type –allele's functions. This usually results an altered molecular
function. Its phenotype is dominant. Exp. Mutation in p53 gene whose
deactivation has a role in cancer.
 Tay-sachs –

Topic 13 – Treatment of genetic diseases

 Modification for the phenotype of genetic disease –

Restriction of substrate uptake (dietary) – PKU
Supplying missing bioactive small molecule (replacement) –thyroid hormone,
congenital hypothyroidism (thyroxine is molecule replaced)
Use alternative metabolic pathway to eliminate toxic metabolite (diversion) - urea
cycle disorders
Use of metabolic inhibitors – inhibitors of cholesterol biosynthesis
Protein therapy – hemophilia A
 Gene therapy - alleviate or overcome disease by introduction of the wild type
gene copy or replacement of the mutant gene with functional gene copy.
Ex vivo – remove tissues or cells alive from bodies to utilize them.
Type Retro viral Adenoviral Adeno-associated
virus
Attribute  Can't be used for  Able to infect wide  need helper virus
s in vivo gene verity of cells to replicate
therapy because  affinity to airway  integrates into
the complement epithelium cornea specific site on
neutralize it. and intestinal chromosome 19
 Infect only epithelium  no risk for
dividing cells  enters cell via insertional
  selectively infects receptor mediated mutagenesis
cancer cells endocytosis  packaging cell line
 Max. Cloning  stays episomal is required for its
capacity is 8kb  used for in vivo production
 not selective for  96% of virus
cancer cells genome was
 disadvantage its deleted
immunogenicity  accommodate
 Max. Cloning inserts up to 4.5
capacity is 35 kb kb

 Vectors in gene therapy –

 Protein/Gene augmentation therapy – in case of loss of function (recessive)

mutations, introduction of the functional gene copy may increase level of the
normal gene product and alleviate or eliminate symptoms. Used for exp. Cystic
fibrosis and Hemophilia A
 Risks in integrating vectors for gene therapy -
1.adenovirus – can elicit an immune response
2.insertional mutagenesis – transferred gene will integrates into the patients
DNA and active proto-oncogene or disrupt tumor-suppressor gene. Leading
possibly to cancer.
3. Insertional inactivation of an essential gene – can occur but will general be
without significant effect cause such a lethal mutations are rare and will kill only
single cells.
 Familial hypercholesterolemia – two strategies
Diversion - of an increased fraction of cholesterol to bile acid synthesis the
single normal LDL receptor gene in these patients can be stimulated to produce
more hepatic receptors for LDL-bound cholesterol.
Inhibition – by drugs that will inhibit enzyme are used to modify the metabolic
abnormalities of inborn errors when the cholesterol load is decreased by
diverting it other compounds or by removing it with physical methods. The liver
tries to compensate for the decreased cholesterol intake by up-regulating
cholesterol synthesis.
 Adenosine deaminase deficiency (ADA) – is one cause of severe combined
immunodeficiency (SCID). The bovine ADA enzyme covalently attached to
polyethylene glycol was administrated by infusion. This treated with enzyme
replacement therapy
 Gaucher disease – it’s a storage disease, inherited as autosomal recessive
caused by deficiency of the enzyme glucocerebroside. The principle of the
treatment is carbohydrates of glycoprotein are modified, terminal sugars are
removed to expose core alpha-mannosyl residues. Enables it to be
phosphorylated and be targeted to the lysosome.
 Stem cell – cell that capable both generating another stem cell and differentiate
to specialized cells within a tissue or an entire organism. But cannot create an
organism. Thus, they pluripotent.
 Nuclear transplantation – transfer of diploid nucleus from an adult donor
somatic cell into oocyte cytoplasm to generate a cloned embryo. This method
generate embryonic stem cells which are genetically identical to donor nucleus.
 So they could be used for cell transplantation to donor without fear of immune
rejection.
 Bone marrow transplantation – is used to treat diseases like
lysosomal storage disease, beta thalassemia and gaucher disease.
 Germ line gene therapy – is banned in all countries
-genetic modification of human gametes, zygotes or the early embryo.
 Somatic cell gene therapy -
-genetic modification of somatic cells and tissues of the patient
-not inherited
-requires permission
 Essential requirements of gene therapy -
-identity of the molecular defect
-a functional copy of the gene
-knowledge of the pathophysiological mechanism
-favorable risk to benefit ratio
-appropriate target cell
-appropriate regulatory components for the transferred gene
 Non-viral ways of gene transfer -
protein DNA conjugates
Naked DNA
artificial chromosome
DNA packaged liposomes
 Gene therapy approaches for cancer -
- direct killing of disease cells- artificial cell killing involves transfer to cells of
gene encoding toxic compounds.
-assisted killing of disease cells by immune system cells – gene of foreign MHC
antigen is evoking an immune response –Cytokine ,TNF these genes have anti-
tumor effect.
-targeted inhibition – inhibition of unwanted responses in autoimmune diseases
 To achieve targeted inhibition we can use -
-antisense oligonucleotides
-siRNA
-ribozyme
they join into disease cells contain the mutant gene that block expression of
pathogenic gene

Topic 14 –Developmental genetics

 Zygote – it’s the fertilized egg, which can develop to an entire organism and
therefore called totipotent.
 Morphogen – it’s a substance produced by cells in particular region of an
embryo that diffuse from point of origin through the tissues to form
concentration gradient cells goes specification and then determination to
different fates developing concentration of morphogen. Helps establish the axes
of the developing embryo according to concentration gradient. Exp. SSH –sonic
hedgehog
 HOX genes – encode transcription factors have 180 base pair sequence known
as the homebox - which encodes 60 amino acid sequence called homedomain
(Helix turn helix)
  Maternal effect – environmental effect that may cause wide variety of birth
defect
maternal effect genes functioning in the mother and needed for development of
the embryo. Correlation to zygotic genes is they affect those gene according to
the info laid out in the egg.
 Specification of cells is reversible | determination of cells isn’t reversible
 Zygotic genes – are genes developmental genes that function in the embryo.
They have correlation to maternal effect genes because they are affected by the
info. Laid out in the egg by the maternal genes.
 Homebox – homeotic genes are transcriptional activators of other genes their
role is to transform the periodicity of embryo into a body plan with liner
differentiation.
Most HOX genes contain Homebox sequence of 180 base pairs. Mutations in
those genes can result in replacement of one body part or segment by another.

Topic 15, 17, 19-20

 Invasive parental tests -

1. Amniocentesis –obtain amniotic fluid which contain cells of fatal origin that
can be cultures for analysis. The fluid is withdrawn from amniotic sac by syringe
from the amnion through the abdominal wall and uterine wall.
2. Chorionic villus sampling (CVS) – biopsy of tissue from the villous area of
the chorion, generally between weeks 10-12.
3. Cordocentesis – obtain sample of fatal blood directly from placenta
4.preimplantion genetic diagnosis- molecular cytogenetic techniques during IVF
to select embryos free of specific genetic condition for transfer to uterus. Can be
performed by micromanipulation to remove polar body or by biopsy of single
cell from 6-8 cell embryo after fertilization.
 Not invasive parental tests-
1.maternal serum alpha fetoprotein – measured in second trimester as screening
for NTD
2.first trimester maternal serum screening – preformed between weeks 11-13
relies on quantify levels of certain substances and measuring substance of lateral
by highly targeted ultrasonography exam. Maternal serum substance are
associated plasma protein A (PAPP-A) and hCG. The PAPP-A is below normal
range in all trisomy and hCG is elevated in trisomy 21 but depressed in the other
ones.
3. Second trimester maternal serum screening – measuring three substances in
mother serum (MSAFP,Free beta-hCG and unconjugated estriol) all of them are
below normal range in all trisomy.
4. Ultrasonography – high frequency soundwaves used to examine internal body
structure.
5. Isolation of fetal cells from maternal circulation.
 Neural tube defect – can be prevented by supplementation of folic acid at least
1 month before conception and continuing through 1st trimester of pregnancy
while decrease the incidence of NTD.
 Fetal sex –is important to know if the parental diagnosis for certain X-linked
recessive may be inherited. Can be checked by invasive tests and ultrasound
exam.
  Disease association – presence or decrease frequency in affected individuals
compared with control.
 Successful Screening for newborn- phenylketonuria , congenital deafness and
congenital hypothyroidism
 Laboratory tests provide basic genetic counseling – karyotype, biochemical
analysis and DNA analysis.
 Ethical principles-
respect for individual autonomy
beneficence
avoidance of maleficence
justice
 Eugenics –increasing prevalence of desirable traits in population by decreasing
the frequency of deleterious allele at relevant loci through controlled selective
breeding. It’s the opposite of dysgenics
 Dysgenics – determination in health and well-being of population by practices
that allow accumulation of deleterious alleles.

Topic 16 – Cancer genetics

 Cancer cells characterized by uncontrolled proliferation

 Consequence of loss of function mutation in p53 – if there isn’t functional
copy of p53 then no DNA damage checkpoint DNA damage is not corrected or
apoptosis isn’t induced cause of the DNA damage.
 Oncogenes – are gain of function mutations of proto-oncogenes which cause
enhance of cell proliferation or inhibit apoptosis.
 Tumor suppressor genes - gene which normally control cell proliferation and
active apoptosis, in loss of function mutation helps cancer progression.
Divided into two groups:
1.gatekeeper- directly suppress tumor formation by negative regulation of cell
proliferation or cell apoptosis. Exp. P53, Rb, p21
2.caretaker – involved in DNA repair and genome maintenance. Exp.BRCA1,
BRCA2
 Retinoblastoma protein – its un-phosphorylated form is attached E2F
transcription factor which prevents its transcriptional activator effect on genes
necessary for DNA synthesis and entering S phase.
 Sporadic cancer – caused by somatic mutation.
 Hereditary cancer – initial cancer causing mutation is inherited in germ line.
 Loss of heterozygosity – when somebody has heterozygous genotype, and one
of his cells normal allele is lost, won't be expressed or become mutated. Its
typical mutation in tumor suppressor genes found in hereditary cancers. Normal
allele can be lost cause of:
1. Deletion
2.duplication
3.insertion
4.frameshift mutation
 Philadelphia chromosome – translocation between 9:22 cause chronic
myelogenous leukemia (CML). ABL gene on chromosome 9 fused to BCR gene
on chromosome 22 results in constitutively active tyrosine kinase.
Topic 18 – Pharmacogenetics

 Metabolic modifications -
phase 1 – hydrolysis/reduction/oxidation/hydroxylation
phase 2- glucronidation/sulphonation/acetylation/methylation. Conjugate with
amino acid
 CYP enzyme – the most important enzyme system in the metabolism is
responsible for activation the drug and conjugating the active secondary
metabolite with glucorinic/sulpharic acid which is followed by excretion.
Those enzymes mainly found in the SER of liver cells but can be found in other
organs i.e. intestinal epithelium. Part of a families as well is highly polymorphic.
 P450 enzyme - the role of it is to eliminate drugs by catalyze the oxidation of
organic substances. Sometime CYP modification of the drug is needed for the
activation of P450. Exp.Terfenadine
if there is a mutation its not visible and its detected by administration of taking
the drug
 Liver – is the most important organ in drug metabolism
SERCYP enzyme drug metabolized
 Sensitivity – if the patient is slow metabolizer of the drug we need lower the
doses because the sensitivity is enhanced. If the patient is fast metabolizer we
need higher doses to keep it in threptic range.
 Contergan – is teratogenicity drug which used to combat nausea in pregnant
women that caused severe thalidomide embryopathies
 Xenobiotics – are drugs, it's an organic chemical which is taken up and found in
organism but not produced normally or found in high concentration in the human
body. Exp. For Xenobiotics – drugs, dioxins, natural compound that been taken
by another organism.
 Succinylcholine – is used as muscular relaxant for intubation anesthesia, it stops
breathing but it's supposed to be hydrolyzed by butyrlcholinesterase enzyme and
recover spontaneous breathing in 1-3 min.
Some people the affinity of the enzyme for the drug is poor and cause prolonged
stop of breathing therefore we need to connect the patient to forced respiratory
machine. Its atypical allele the trait is autosomal recessive the frequency is
1/2000
 Environmental xenobiotics interfere with drug metabolism – yes, there is a
link between the two (Eco-genetics and pharmacogenetics). Exp. Hemolytic
shock in G6PD deficiency can evolved by drugs and some natural compounds
present in our environment (cause of beans for exp.)
 Diet may effect on drugs -
grapefruit is an inhibitor of enzymes
Cabbage, broccoli are enzyme inducers