Enhanced Diagnostic Tools
Micro RNAs (miRNAs) are molecules synthesized in the nucleus, (more precisely small noncoding RNA’s) as long primary transcripts (pri-miRNA); this transcript is cleaved in the so called microprocessor complex, (made up of the enzymes Drosha and Pasha) into an stem-loop structure of about 70 bp’s(pre-miRNA).
The pre-miRNA, then moves into the cytoplasm, where the RNA -induced silencing complex (RISC) is activated, and cuts the pre-miRNA into 20-25 nt’s, which subsequently targets the corresponding mRNA, which is then cleaved and inactivated.
Microprocessor Complex. Refers to the enzyme complex, where the major components are the enzyme called Drosha ( a class II RNAse III enzyme) and Pasha (a double stranded RNA binding protein, also known as DGCR8). This complex converts the primary RNA transcript (pri-miRNA) into a 70 bp’s mi-RNA (pre-miRNA).
Dicer is a RNase III endonuclease enzyme that cleaves double-stranded RNA (dsRNA) and pre-microRNA (miRNA) into short double-stranded RNA fragments called small interfering RNA (siRNA) of about 22 nucleotides long, that have two-base overhang on the ends. Dicer initiates the first step in the RNA interference (RNAi) pathway and is part of the RNA-induced silencing complex (RISC), whose catalytic component Argonaute is an endonuclease able of degrading messenger RNA (mRNA) whose sequence is complementary to that of the siRNA guide strand.Therefore Dicer plays a role in RISC by providing the initial RNA material to activate the complex and also the first RNA substrate molecule. When Dicer cleaves dsRNA and pre-miRNAs, a 20-25 base pair double-stranded RNA fragment is formed.
RNA-Induced Silencing Complex (RISC). Refers to the multi-protein aggregate, where the RNA's enzyme Dicer is the principal endonuclease and cleaves dsRNA and binds short antisense RNA strands (guide strands) which then bind complementary strands. Upon binding of the complementary strand, it activates RNase activity and cleaves the RNA. This mechanism is important in both gene expression control by microRNAs and to fend off viral infections, where double-stranded RNA is frequently the infectious vector.
Diagram of the Micro RNA Process
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