RNA INTERFERNCE OR

POST TRANSCRIPTIONAL
GENE SILENCING
GROUP MEMBERS
 ANUM REHMAN
 AIMEN WAHEED
 ZARMEENA
 SARA SALIM
 HUZEMA MUNIR
CONTEXT
 INTRODUCTION
 PROCESS AND MECHANISM
 APPLICATIONS
INTRODUCTION
 It is also called as co-suppression or quelling.
 The conserved biological process in which RNA
molecules inhibit gene expression or can direct
repression of genes with homology to those short
RNAs.
 This repression, is called as RNA interference
(RNAi).
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 RNA interference can inhibit the gene
expression in three possible ways:-
• Destruction of the mRNA.
• Translational inhibition of the mRNA.
• Transcriptional silencing of the promoter that
directs expression of that mRNA.
.
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 The RNAi pathway is found in many eukaryotes
including animals.
 Initiated by enzyme DICER.
 Which cleave long dsRNA into short double
stranded fragments of approx 20 nucleotides
siRNA.
 Two types of small RNA molecules – microRNA
(miRNA) and small interfering RNA (siRNA) – are
central to RNA interference.
PROCESS AND MECHANISM
 RNAi is an RNA-dependent gene silencing process that
is contolled by the RNA-induced silencing complex
(RISC) and can be initiated either by:-
 Short interfering RNAs (siRNAs):-
• They are produced from dsRNA by the help of an
enzyme Dicer (ribonuclease protein).
• The products of this are short dsRNA fragments about
23 nucleotides long (short interfering RNAs) in the
cytoplasm of a cell.
• They interact with the catalytic RISC component
argonaute (active protein component of RISC ).
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 Exogenous dsRNA:-
• When the dsRNA is exogenous (coming from
infection by a virus with an RNA genome), the RNA
is imported directly into the cytoplasm and cleaved
to short fragments by Dicer.
 Endogenous dsRNA:-
• The initiating dsRNA can also be endogenous
(originating in the cells).
• The primary transcripts from such genes are first
processed to form the stem loop structure of
miRNA in the nucleus, then exported to the
cytoplasm.
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 MicroRNAs (miRNAs):-
• They are naturally occuring RNAs that directs the
repression of genes in the same way as siRNAs.
• They are 21or 22 nts long, arise from larger
precursors (about 70-90 nts long) transcribed from
non-protein encoding genes.
• These transcripts contain sequences that form stem
loop structures, which are processed by Dicer.
STEPS OF RNAi
 The interference is due to the presence of
antisense RNA which is complementary to the
specific cellular mRNA.
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 The complementarity between the antisense
strand RNA and the cellular mRNA causes the two
molecules to bind to each other.
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 The current RNA interference involve two steps:
 Creation of an RNA induced silencing complex
(RISC)
 The degradation of cellular mRNA by the
endonucleases component of RISC known as
argonaute.
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 CREATION OF RISC COMPLEX OCCURS WHEN
 Double stranded RNA molecules are present in the
cell.
 These dsRNAs are then recognized by the
enzyme called as DICER.
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 That cut them into pieces that are 21 to 23 nts
long.
 These short dsRNAs formed are called as siRNA
(shorter units).
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 An antisense strand of the siRNAs then associates
with the catalytic component of RISC called as
argonaute.
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 Then that complex associates with the cellular
mRNA that has the region of sequencing homology
with the RNA fragments in the RISC.
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 Then mRNA is degraded.
 This can reduce and even eliminate the translation
of the messenger RNA for the gene.
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 APPLICATIONS…
 Medicines and therapy:-
 RNA interference is also exploited in therapy.
 They were first used in the treatment of macular
degeneration (medical condition which results in loss of
vision because of damage to the retina) and respiratory
syncytial virus ‘RSV’ (a virus that causes respiratory tract
infections).
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 Cancer:-
RNA interference is also used for the treatment of
cancer by silencing genes :-
 Differentially upregulated in tumor cells.
 Or genes involved in cell division.
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 Gene knockdown:-
The RNAi pathway is often used in experimental
biology to study the function of genes in cell culture.
 Using this mechanism, researchers can cause a
drastic decrease in the expression of a targeted
gene.
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 Biotechnology:-
RNA interference has been used for applications in
biotechnology e.g :

 Food:-
RNAi has been used to genetically engineer plants
to produce lower levels of natural plant toxins e.g in
cotton seeds.
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 Transgenic plants:-
Transgenic crops have been made to express small
bits of RNA, carefully chosen to silence crucial
genes in target pests.
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 Antiviral:-
 RNA interference is also used to treat infections
caused by herpes simplex virus type 2 and the
inhibition of viral gene expression in cancerous
cells.