miRNA Processing and its

Amplification
MIC 4203 CE1 (T2) Class Presentation
Name:- Rohit Shirish Mali
Roll No.:- 353
Introduction
• MicroRNAs (miRNAs) are small, regulatory, noncoding RNA
molecules that control the expression of their target mRNAs.
• The functional form of an miRNA is typically ~21 to 22 nucleotides
long.
• miRNAs are generated either from the processing of a host intron or
by transcription from their own dedicated promoters.
• Synthesized by RNA Pol II and some are synthesized using RNA Pol
III.
• The miRNAs are produced as large RNA primary transcripts called
pri-miRNA that are self-complementary.
• miRNAs mainly exert their function post-transcriptionally by
modulating mRNA stability and/or protein translation.
Processing of miRNAs
Processing of miRNA takes place by two distinct pathways:-
A. Canonical pathway
B. Non canonical pathway

C. Canonical pathway
• This pathway consists of 4 steps:-
i. Pri-miRNA biogenesis
ii. Processing from pri-miRNA to pre-miRNA
iii. Exporting pre-miRNA into cytoplasm
iv. Mature miRNA generation
i. Pri-miRNA biogenesis:-
• Stem loop structured pri-miRNA is synthesized from genomic DNA
by RNA Pol II

Fig. Structure of pri-miRNA

Image source:-James D. Watson, Tania Baker, Stephen P. Bell, Alexander Gann,

Michael Levine, Richard Loswick (2014) Molecular Biology of the Gene, 7th
Edition, Pearson Education, Inc. and Cold Spring Harbor Laboratory Press, Inc.
ii. Processing from pri-
Fig. Recognition
miRNA to pre-miRNA:- and cleavage of
• Pri-miRNA is cleaved by pri-miRNA by
microprocessor
Drosha: an Rnase III like complex
enzyme that recognizes and
digests stem-loop structures
formed in pri-miRNA. Image source:-
• Drosha works together with James D. Watson,
Tania Baker, Stephen
another protein subunit (called P. Bell, Alexander
Pasha in some organisms and Gann, Michael
Levine, Richard
DGCR8 in others) forming a Loswick (2014)
microprocessor complex. Molecular Biology of
the Gene, 7th
• For processing by Drosha 3’ Edition, Pearson
Education, Inc. and
and 5’ flanking regions are Cold Spring Harbor
needed. Laboratory Press,
Inc.
iii. Exporting pre-miRNA into cytoplasm:-
• Pre-miRNA is exported to cytoplasm by Exportin-5.
• This is rate determining step in miRNA processing.

iv. Mature miRNA generation:-

• Pre-miRNA is processed by Dicer producing a ~22 nucleotide
miRNA.
• Dicer is composed of 3 domains: 2 RNase III domains and a ds-RNA
binding domain called PAZ domain.
B. Non Canonical Pathway:-
• Subsets of intronic miRNA are “mirtron” whose processing is
independent of Drosha.
• The mirtrons lack the lower hairpin and the flanking single-stranded
regions that recruit the microprocessor complex.
• Instead, they contain 5' and 3' splice sites that are presumably
recognized by small nuclear RNAs (snRNA).

Fig. Non canonical pathway of miRNA processing

Image source:-Hawley, Zachary C. E. & Campos-Melo,
Danae & Droppelmann, Cristian & Strong, Michael. (2017).
MotomiRs: miRNAs in Motor Neuron Function and
Disease. Frontiers in Molecular Neuroscience. 10.
10.3389/fnmol.2017.00127.
Image source:-Shukla
GC, Singh J, Barik S.
MicroRNAs: Processing,
Maturation, Target
Recognition and
Regulatory Functions. Mol
Cell Pharmacol.
2011;3(3):83-92. Fig. Processing of miRNA
Amplification of miRNA
• RNAi silencing is extremely efficient because
very small amount of dsRNA are often enough
to induce a complete shutdown of target gene
expression
• Amplification is the major factor contributing
to this efficiency.
• RNA-dependent RNA polymerase(RdRP) is
enzyme required in this process
• RdRP can amplify inhibitory signal: The Fig. Amplification of miRNA
RdRP generates dsRNA after recruitment to the Image source:-James D. Watson, Tania
mRNA by original miRNA Baker, Stephen P. Bell, Alexander Gann,
Michael Levine, Richard Loswick (2014)
• Amplification is of major importance in Molecular Biology of the Gene, 7th
Edition, Pearson Education, Inc. and Cold
centromere silencing Spring Harbor Laboratory Press, Inc.
References
• Shukla GC, Singh J, Barik S. MicroRNAs: Processing, Maturation,
Target Recognition and Regulatory Functions. Mol Cell Pharmacol.
2011;3(3):83-92.
• Jiang S, Yan W. Current View of microRNA Processing. Signal
Transduction Insights. January 2016. doi:10.4137/STI.S12317
• James D. Watson, Tania Baker, Stephen P. Bell, Alexander Gann,
Michael Levine, Richard Loswick (2014) Molecular Biology of the
Gene, 7th Edition, Pearson Education, Inc. and Cold Spring Harbor
Laboratory Press, Inc.
• Lewin’s Genes XI, (2014) Jones and Bartelett Publishers Inc.
• W.S. Klug and M.R. Cummings, Concepts of Genetics,12th edition
(2019) Pearson education
THANK YOU