Gene Silencing

Niranjana.S.Karandikar MSc-I Roll No.12

Contents
Introduction
Cellular component of GS Level of GS

Transcriptional GS(TGS) Post-Transcriptional GS(PTGS)

Applications

Introduction
"switching off" of a gene by a mechanism other than genetic modification. A gene which would be expressed ("turned on") under normal circumstances is switched off by machinery in the cell. Gene silencing occurs when RNA is unable to make a protein during translation. Partial or complete inactivation of gene is known as Gene Silencing.

Cellular Components Of Gene Silencing

Histone Chromatin microRNA Small interfering RNA Double stranded RNA piRNA Transposable element Dicer

Histones
Highly alkaline proteins Found in eukaryotic cell nuclei Package and order the DNA into structural units called nucleosomes. Chief protein components of chromatin. Act as spools around which DNA winds.

Chromatin
Combination of DNA and proteins. The primary protein componentshistones. DNA coding genes actively transcribed ("turned on") more loosely packaged found associated with RNA polymerases = euchromatin. DNA coding inactive genes ("turned off") found associated with structural proteinsmore tightly packaged=heterochromatin

Micro RNA
abbreviated as miRNA. A microRNA molecule has very few nucleotides (an average of 22). miRNAs are post-transcriptional regulators. Binds to complementary sequences on target mRNAs, resulting in translational repression or target degradation and gene silencing.

Small interfering RNA

Abbreviated as siRNA. Sometimes also k/as short interfering RNA or silencing RNA. It is a class of dsRNA molecules. 20-25 nucleotides in length. Role of siRNA is its involvement in the RNA interference (RNAi) pathway, where it interferes with the expression of a specific gene.

Double stranded RNA

RNA with two complementary strands Similar to the DNA found in all cells. Can trigger RNA interference in eukaryotes.

piRNA
Piwi-interacting RNA : RNA-protein complexes It is the largest class of small non-coding RNA molecules that is expressed in animal cells. piRNAs form with piwi proteins. through interactions

Transposable Element (TE)

DNA sequence that can change its relative position (self-transpose) within the genome of a single cell. Transposition can create phenotypically significant mutations and alter the cell's genome size.

Dicers
Endoribonuclease in the RNase III family. It cleaves (dsRNA) and pre-miRNA into short double-stranded RNA fragments called siRNA.

Levels Of Gene Silencing

It occurs at two levels : Transcriptional Gene Silencing-TGS. Post transcriptional Gene Silencing- PTGS.

However sometimes it occurs during meiosis.

Transcriptional Gene Silencing:

a) Genomic Imprinting b) Paramutation c) position effect d) RNA-Directed DNA Methylation e) Transposon silencing f) Transgene silencing g) Transcriptional gene silencing

Genomic Imprinting
genomic imprinting is a genetic phenomenon by which certain genes are expressed in a parent-of-origin-specific manner. It is an inheritance process independent of the classical Mendelian inheritance. Imprinted genes are either expressed only from the allele inherited from the mother (eg. H19 or CDKN1C), or in other instances from the allele inherited from the father (eg. IGF-2). Forms of genomic imprinting have been demonstrated in insects, mammals and flowering plants.

Post-transcriptional Gene Silencing

a) RNA interference b) Nonsense mediated decay

RNA Interference
RNA interference (RNAi) is a system within living cells that helps to control which genes are active and how active they are.Two types of small RNA molecules microRNA (miRNA) and Small interfering RNA (siRNA) Are central to RNA interference. RNAs are the direct products of genes, and these small RNAs can bind to specific other RNAs and either increase or decrease their activity, for example by preventing a messenger RNA from producing a protein. RNA interference has an important role in defending cells against parasitic genes viruses and transposons but also in directing development as well as gene expression in general.

Difference between TGS & PTGS

TGS Promoters silenced Genes hypermethylated in promoter region. Prevent Transcription. PTGS Promoters active Genes hypermethylated in coding region. Prevent Translation.

Significance
Supression of undesired characters Silencing of Onco-genes In-vitro;In-vivo silencing Correctional method Vaccinations

References:
http://en.wikipedia.org/wiki/Gene_silencin g http://www.wisegeek.com/what-is-rnainterference.htm http://www.wisegeek.com/what-is-genesilencing.htm http://www.sciencedaily.com/releases/200 6/10/061002152958.htm

Thank YOu