GENE SILENCING

By: Sajjad Ahmad

PML02141003

Definition

Gene silencing is a general term used to

describe the epigenetic regulation of gene
expression.
In particular, this term refers to the ability
of a cell to prevent the expression of a

certain gene.

Cellular components
Histones

Chromatin and heterochromatin

miRNA
siRNA
dsRNA
Dicer
Transposons

Material covered:

Transcriptional gene silencing.

Post transcriptional gene silencing.
Meiotic gene silencing

Its applications in research.

 Gene silencing can occur during either

transcription or translation.

Often used in research. (To produce

therapeutics)

 Do not confuse gene silencing with gene

knockout.

 In gene silencing the expression of a

gene or genes are reduced.

Whereas
In gene knockout the whole gene is

eradicated from the genome thereby

having no expression at all.

Types of gene silencing

1. Transcriptional gene silencing.

Includes:

Paramutation.

Transposon silencing.

Position effect.

2. Post transcriptional gene silencing.

Includes:

RNA interference.
Nonsense mediated decay.

3. Meiotic gene silencing

Transvection

1. Transcriptional Gene Silencing

Result from histone modifications or DNA
methylation creating an environment of

heterochromatin around a gene that makes

it inaccessible to transcriptional machinery

(RNA polymerase, transcription factors,

etc.).

Paramutation:

A paramutation is an interaction between

two alleles at a single locus, whereby one

allele induces a heritable change in the other
allele.

The allele inducing the change is said to be

paramutagenic, whilst the allele that has
been epigenetically altered is termed

paramutant (or paramutated).

contd:
A paramutant allele may have altered levels of gene

expression,

May continue in offspring which inherit that allele, even

though the paramutagenic allele may no longer be

present.

Occurs mostly in plants.

Through proper breeding, paramutation can result in

sibling plants that have the same genetic sequence, but

with drastically different phenotypes.

Transposon silencing.

A Transposon is a DNA sequence that can change its

position within the genome, sometimes creating or

reversing mutations and altering the cell's genome size.
Transposition often results in duplication of the TE.
Barbara McClintock's discovery of these jumping

genes earned her a Nobel prize in 1983.

Contd:
Transposon silencing is a form of transcriptional

gene silencing targeting transposons.

The jumping of transposons have the tendency to

generate genomic instability and can cause

extremely deleterious mutations.
Transposable element insertions have been linked to

many diseases including hemophilia, severe

combined immunodeficiency etc.

Contd:
Silencing of transposons is therefore very
important in the germ line in order to stop

transposon mutations from developing and

passing on to next generations.

Studies show that siRNA play a major role

in silencing of transposons.

Position effect:

Position effect is the effect on the expression of

a gene when its location in a chromosome is changed,

often by translocation.
This has been well described in Drosophila with respect

to eye color and is known as position effect

variegation (PEV)

Post-transcriptional gene silencing

Post-transcriptional gene silencing is the result of

mRNA of a particular gene being destroyed or blocked.

The destruction of the mRNA prevents translation to

form an active gene product in most cases, a protein.

A common mechanism of post-transcriptional gene

silencing is RNAi.

RNA interference

The phenomenon of inhibition of gene expression by

RNA molecules is called RNA interference (RNAi)

RNAi is caused by small interfering RNAs (siRNAs)

RNA interference:

RNA interference (RNAi) is a natural process used

by cells to regulate gene expression.

discovered in 1998 by Andrew Fire and Craig Mello,

who won the Nobel Prize for their discovery in

2006.

 The process to silence genes first begins with the entrance

of a double-stranded RNA (dsRNA) molecule into the cell.

This triggers the RNAi pathway.
The double-stranded molecule is then cut into small

double-stranded fragments by an enzyme called Dicer.

These small fragments, which include small interfering

RNAs (siRNA) and microRNA (miRNA), are

approximately 21-23 nucleotides in length.

 The fragments integrate into a multi-subunit

protein called the RNAi induced silencing complex

(RISC), which contains Argonaute proteins that
are essential components of the RNAi pathway.
There are 2 strands of the fragment (guide strand)

and (Passenger strand).

The guide strand binds to RISC whereas
The passenger strand is degraded.

 The guide strand of the fragment remains bound to

RISC and directs the sequence-specific silencing of

the target mRNA molecule.
siRNA molecules have the ability to silence genes

bye causing the endonucleatic cleavage of the target

mRNA molecules.
miRNA molecules have the ability to suppress

translation of the mRNA molecule.

RNAi is thought to have evolved as a cellular

defense mechanism against invaders, such
as RNA viruses, or to combat the proliferation
of transposons within a cells DNA.

RNA Interference

Meiotic gene silencing

Transvection:
phenomenon,
results
from
an
interaction between an allele on one chromosome
and the corresponding allele on the homologous
chromosome.
Transvection can lead to either gene activation or
repression.
Formally, it can also occur between non allelic
regions of the genome as well as regions of the
genome that are not transcribed.
Epigenetic

Nonsense-mediated mRNA decay

(NMD)
Its main function is to reduce errors in

gene expression by eliminating mRNA

transcripts that contain premature stop
codon.

If these aberrant mRNA transcripts were

translated, the result would be deleterious

gain-of-function or dominant-negative
activity of the resulting proteins.

Applications:

Gene silencing techniques have been widely used by

researchers to study genes associated with

disorders. These disorders include:
cancer,
infectious diseases,
respiratory diseases,
and neurodegenerative disorders.

Thank you