25-12-2015

T. Gayathri Devi

Genotype
Phenotype
Chargaffs rule ( Purine= pyrimidine)
Exons / Introns
Allele ( Eg- Rh D, d)
Gene

AA
(Homozygous)

aa
(Homozygous)

Aa
(Heterozygous)

Net expression

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Do not encode proteins

Inhibit gene expressions
1000 genes in human genome encode mi
RNA
si RNA ( small interfering RNA) introduced
by investigator into cell

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Genetic disorders

single gene
disorder
Mendelian
inheritance

Non
mendelian
inheritance

Permanent change in DNA

Chromosomal
disorders

Multifactorial
disorders

Numerical
disorders

Structural
disorders

>1 mutant
gene+
environment

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

25-12-2015

Silent mutation

Missense mutation

Non sense mutation

Dr. T.Gayathri Devi

Eg- Cystic fibrosis Deletion of

phenylalanine at 508th position in CFTR
gene
Dr. T.Gayathri Devi

Mechanism
Variable Expression

Description
Most genetic diseases vary in the
degree of phenotypic expression:

Incomplete penetrance

when some individuals who have the

disease genotype do not display the
disease phenotype

Delayed age of onset

Many individuals who carry a

disease-causing mutation do not
manifest the phenotype until later in
life.
Pleiotropy exists when a single
disease-causing mutation affects
multiple organ systems.

Pleiotrophy

Locus heterogeneity

6th position of chain

Hb Milwaukee

Glutamate

Hb Bristol

Aspartate

Hb Sydney

Alanine

Acceptable mutation

Hb hikari ( chain )

Partially acceptable
mutation

Hb S ( chain )

Unacceptable
mutation

Hb M Boston (
chain )

Coding codon Stop

codon

-thalassemia

Stop codon Coding

codon

-thalassemia

Dr. T.Gayathri Devi

Hemoglobin

Trinucleotide repeat sequence

Normal number of repeats- 29
Dynamic mutation
Eg Coding region- Huntingtons chorea
Non coding region - Fragile X syndrome
Splice site mutation Myotonic dystrophy , thalassemia

Dr. T.Gayathri Devi

1. autosomal recessive
2. autosomal dominant (>50% of all single
gene dz)
3. X-linked dominant
4. X-linked recessive

Locus heterogeneity exists when the

same disease phenotype can be
caused by mutations in different loci
Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

25-12-2015

Most common
Expressed in
heterozygous state

Expressed only in
homozygous state
Uniparental disomy
( when one parent
is carrier and other
normal- both gene
received from
carrier parent)

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Mechanisms of Generating UPD

Normal

Nondisjunction in one
Nondisjunction parent, followed by
in both parents duplication in embryo

Gamete

Zygote

Duplication

Embryo

(a)

(b)

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

(c)

Both alleles are expressed

Eg Blood group antigens A and B

Gain of function
mutation

New protein synthesized

less common
Eg huntingtons disease

Loss of function
mutation

Huntingtin synthesized
Toxic to neurons

Dr. T.Gayathri Devi

Reduced protein or
defective protein
synthesis
More common
Eg. Osteogenesis
imperfecta
Some are Dominant
negative mutant allele

Dr. T.Gayathri Devi

25-12-2015

Dominant negative mutant allele are mostly

ass with loss of function mutations
Eg. One defective collagen chain in the
trimer affects collagen helix assembly

System
Huntington disease
Neurofibromatosis
Myotonic dystrophy
Tuberous sclerosis

Urinary

Polycystic kidney disease

Gastrointestinal

Familial polyposis coli

Hematopoietic

Hereditary spherocytosis
von Willebrand disease

Skeletal

Marfan syndrome
Ehlers Danlos syndrome (some variants)
Osteogenesis imperfecta
Achondroplasia

Metabolic

Familial hypercholesterolemia
Acute intermittent porphyria

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Mutation of fibrillin gene in 15q21

Affects elastic fibres
Cause of death Aortic dissection ( so treat
HT)

Dr. T.Gayathri Devi

NF 1 ( Von
Recklinghausen )

Disorder

Nervous

More common (90%)

NF 1 gene ( ch 17)
neurofibromin
Neurofibromas
6 or more cafau lait
spots
Lisch nodules in iris
Scoliosis, bone cysts
risk of meningiomas,
phaeochromocytoma,
wilms tumour

NF 2 ( BL acoustic
neurofibromatosis

Less common
NF 2 gene ( ch 22)
merlin
Bilateral acoustic
neuromas
Multiple meningiomas
Caf au lait spots
No lisch nodules

Juvelile myelomonocytic
leukemia is commonest
leukemia
in children
with NF 1
Dr. T.Gayathri
Devi

Dr. T.Gayathri Devi

Defect in collagen synthesis

Heterogenous group of disorders
Different modes of inheritance (AR/AD)
Hyperextensible skin ( cigarette paper skin)
Hyperextensible joints
MC Hypermobility type (AD)
MC AR- Kyphoscoliosis
Most dangerous- Vascular type

Dr. T.Gayathri Devi

25-12-2015

System
Nervous

Metabolic

Hematopoietic
Endocrine
Skeletal
Dr. T.Gayathri Devi

Disorder
Neurogenic muscular atrophies
Friedreich ataxia
Spinal muscular atrophy
Cystic fibrosis
Phenylketonuria
Galactosemia
Homocystinuria
Lysosomal storage disorder
-Antitrypsin deficiency
Wilson disease
Hemochromatosis
Glycogen storage diseases
Sickle cell anemia
Thalassemias
Congenital adrenal hyperplasia
Ehlers Danlos syndrome (some variants)
Alkaptonuria
Dr. T.Gayathri Devi

Def of enzyme phenylalanine hydroxylase

Light colour
hair & skin
Mousy/must
y odour
Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

X linked
recessive

Andersons
Ma Ardles

Pompes

Forbes

- Genetic def of
functional
lysosomal
enzymes
- RE system most
affected as
abundant in
macrophages

Von gierkes

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

25-12-2015

Common in ashkenazi jews

Normal at birth
Also cherry red spot

Neurons- micro

TYPE- A
Severe infantile form
HS with CNS invol
Death within 3 yrs

EM- Zebra bodies

TYPE- B
Organomegaly
without CNS invol

Dr. T.Gayathri Devi

Mutation in hexosaminidase enzyme ch 15

TYPE- C
More common

Dr. T.Gayathri Devi

excess lysosomal GM 2 ganglioside accu

Neuronal ballooning
Cherry red spot in macula (lipid in retinal
cells)
EM- lysosomes onion skin appearance

Neurons most severly affected

Motor/mental deterioration & blindness
( 6 months)
( 2-3 yrs)

Dr. T.Gayathri Devi

MC lysosomal storage disorder

Glucocerebrosidase def
Acc of glucocerebroside in lysosomes of RE
system

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Hepatospleenomegaly
Hypersplenism
Lymphadenopathy
BM involvement

Crumpled tissue
paper cytoplasm

Dr. T.Gayathri Devi

25-12-2015

System

Duchenne muscular dystrophy

Menkes disease
Pelizaeus Merzbacher disease

Hematopoietic

Hemophilia A and B
Chronic granulomatous disease
G6PD deficiency

Immune

Agammaglobulinemia
Wiskott-Aldrich syndrome

Metabolic

Diabetes insipidus
Lesch-Nyhan syndrome

Nervous

Fragile X syndrome

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Occ females may show symptoms due to

random inactivation of X chromosome

Eg. G6PD deficiency women may show

symptoms of varying severity
Dr. T.Gayathri Devi

Disease

Musculoskeletal

Vitamin D resistant rickets.

Incontinentia pigmenti.
Oro facial digital syndrome

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Mitochondrial inheritance
Genomic imprinting
Triple repeat mutations
Gonadal /germline mutation

Dr. T.Gayathri Devi

25-12-2015

Maternal inheritance (only ova had

mitochondria)
Sons do not transmit
disease

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Organs- containing large mitochondrial

content- CNS, Skeletal muscle, cardiac
muscle, liver, kidney
Egs- Lebers optic neuropathy, Leighs
disease, mitochondrial encephalopathy lactic
acidosis and stroke like syndrome (MELAS),
NARP syndrome

Dr. T.Gayathri Devi

Maternal imprinting silencing of

maternal gene
Paternal imprinting silencing of
paternal gene
If expressed gene deleted ---- disease
If imprinted gene deleted---- no effect
Eg- Praderwilli syndrome & Angelman
syndrome
Beckwith Wiedmann syndrome &
Albrights hereditary osteodystrophy
Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Deletion of paternal
chromosome which
is usually
expressed while
maternal ch is
imprinted
MR, obesity,
hypogonadism,
hypotonia

Dr. T.Gayathri Devi

25-12-2015

Deletion on
maternal ch 15
MR, seizures,
ataxia,
inappropriate
laughter ( Happy
puppets)
Other cause
uniparental disomy

Dr. T.Gayathri Devi

X linked recessive
CGG repeats
FMR 1 gene affected ( chr appear broken in
karyotyping)
2nd common cause of MR ( 1st- downs)
Long face, large mandible, large ears and
large testicles

Expansion in coding region

Expansion in non coding region

Loss of function

Gain of function

Mutant proteins aggregates

intranuclear inclusions

Mutant protein interferes with

other protein

Fragile X
Friedrich ataxia
Myotonic dystrophy

Huntingtons chorea
Spinocerebellar ataxia
Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Long repeating sequence of 3 nucleotides

Dynamic
Amplification occurs during gametogenesis

Normal 10 to 55 repeats
Premutation 55 200 repeats
Mutation 200 to 4000 repeats
Amplification occurs during oogenesis
Sherman paradox ( Risk of MR greater in
grandson than in brother of transmitting
male)
Anticipation ( worsening of CF in successive
generation)
Diagnosis PCR / Genetic counseling
Southern blot
Dr. T.Gayathri Devi

CAG repeats in ch 4
AD inheritance
Early progressive dementia, choriform
movements

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

25-12-2015

Non dysjunction
happens during
mitotic division that
occurs after fusion of
zygote
Eg- Turner mosaicOne cell gets extra
ch (47, XXX) while
other gets 45,X
So individual has
different cell types

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Study of chromosomes
Metaphasic arrest with colchicine (Mitotic)
Cells- Skin fibroblast, Blood lymphocytes,
amniotic cells
Chromosomes stained
Arranged big to small (descending order)
Usually diploid (46XY or 46XX)

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

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25-12-2015

Dr. T.Gayathri Devi

Dye used

Dr. T.Gayathri Devi

Q Banding

G Banding

R Bandin
(Reverse)

C banding

Quinacrine
mustard

Trypsin foll
by Giemsa

Alkaline sol Chemical foll

foll by
by Giemsa
Giemsa

Microscope
used

Fluoroscenc Light
e
microscopy

Light

Light

Special
Feature

Temporary
Not used

Pattern opp
to G
banding
Rearrange
ment of
terminal
ends

Chromosomal
translocation
invol
centromeric
regions

Permanent
MC

Dr. T.Gayathri Devi

T- banding- for telomeres

C- banding- For constitutive heterochromatin
NOR banding- For nucleolar reorganising
region
FUdR banding- for identifying fragile sites on
chromosomes

Dr. T.Gayathri Devi

X chr

Y chr

Not seen
in humans

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

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25-12-2015

AUTOSOMES

TRISOMY-21 (DOWN SYNDROME)

8, 9, 13 (Patau), 18 (Edwards), 22
22q.11.2 deletion

N+
1

SEX

N1

When fuses with normal gametes

2n+1
Trisomy

2n-1
Monosom
Dr. T.Gayathri Devi
y

Dr. T.Gayathri Devi

Most trisomies (monosomies, aneuploidy) are from

maternal non-disjunction
(non-disjunction or anaphase lag are BOTH
possible)

#1

Dr. T.Gayathri Devi

a. Karyotype: 47 XX or XY+21
b. Most common of the chromosomal disorders
c. Most common cause of inherited mental
retardation
d. Incidence: 1in 700 births
e. Risk increases with maternal age
f. Pathogenesis
i. Meiotic nondisjunction (95%)
ii. Robertsonian translocation (4%)
iii. Mosaicism due to mitotic nondisjunction
during embryogenesis (1%)

Dr. T.Gayathri Devi

CHROMOSOMES

KLINEFELTER: XXY, XXXY, etc.

TURNER: XO

cause of mental retardation

Maternal age related

Congenital Heart Defects, risk for acute leukemias,
GI atresias

Dr. T.Gayathri Devi

Clinical findings
i. Severe mental retardation
ii. Mongoloid facial features (flat face, lowridged nose, and epicanthal folds)
iii. Brushfield spots-speckled appearance of
the iris
iv. Muscular hypotonia
v. Broad short neck
vi. Palmar (simian) crease
vii. Congenital heart defects - endocardial
cushion defect
viii. Duodenal atresia ("double-bubble" sign)
ix. Hirschsprungs disease
x. Increased risk (IS-20x) of ALL
xi. Alzheimer disease (by age 40 virtually all
will develop Alzheimer
Dr. T.Gayathri Devi

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25-12-2015

Cri du Chat syndrome

a. Karyotype: 46 XX or XY, 5p-.
b. Pathogenesis: deletion of the short arm of
chromosome 5
c. Clinical findings
i. Characteristic high pitched cat-like cry
ii. Mental retardation
iii. Congenital heart disease
iv. Microcephaly

Dr. T.Gayathri Devi

a. Karyotype: 47 XXY
b. Caused by meiotic
nondisjunction
c. Common cause of

male hypogonadism

d. Lab diagnosis
i. Elevated FSH and
LH
ii. Low levels of
testosterone
Dr. T.Gayathri Devi

CVS- Mitral
valve prolapse

Dr. T.Gayathri Devi

a. Karyotype: 45XO
b. Common cause of female hypogonadism
c. The second X chromosome is necessary for
oogenesis and normal development of the ovary
d. No Barr body present
e. MC sex chromosome abnormality in females
. Clinical features
i.Failure to develop secondary sex characteristics

Dr. T.Gayathri Devi

-Congenital heart
disease coarctation of the
aorta, bicuspid
aortic valve
- Hydrops fetalis
- mosaicism are at
risk for
gonadoblastoma

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

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25-12-2015

Polymerase chain reaction

Exponential amplification of DNA
Few DNA copies
DNA polymerase and
thermal cycling
Large number of DNA
copies
abnormal sequence easily
identified

Direct sequencing
Indirect methods- Allele specific PCR
RFLP
Genome wide analysis- to identify variants
with low penetrance and small effects

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

To identify
structural
chr
aberrations
in cancer
cells and
other
disease

Fluorescent In-

Situ
Hybridization

Uses fluorescent labelled

DNA fragments, ~10,000
base pairs, to bind (or not
bind) to its complement

Dr. T.Gayathri Devi

SUBTLE

MICRODELETIONS
COMPLEX TRANSLOCATIONS
AND TELOMERE ALTERATIONS

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Check for mitochondrial inheritance

Check if dominant or recessive

X linked or autosomal

Dr. T.Gayathri Devi

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25-12-2015

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

Dr. T.Gayathri Devi

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