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Griffin:siRNA transfection: Difference between revisions
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[[Image:siRNA flow chart.jpg|thumb|right|RNAi pathway]] | [[Image:siRNA flow chart.jpg|thumb|right|RNAi pathway]] | ||
[http://en.wikipedia.org/wiki/RNA_interference RNA interference] is an epigenetic system within living cells that modulates | [http://en.wikipedia.org/wiki/RNA_interference RNA interference] is an epigenetic system within living cells that modulates global gene expression fluctuations. | ||
The native RNAi pathway involves a grooming process where long double-stranded RNA (dsRNA) molecules are refined into double stranded RNA of ~20 nucleotides (first step in the figure to the right). However, the ~20 nucleotide double strand RNA (a substrate for Dicer/R2D2) is now a conventional research product known as siRNA. These ~20 nucleotide siRNA inside a cell is catalytic unwound and one of the two strands, known as the guide strand or antisense strand, is then loaded into the RNA-induced silencing complex (RISC). Experimentally, any mRNA that shares 100% sense compliment sequence to the 'loaded' RISC complex, will bind and undergo nuclease-dependent degradation. As mRNA levels go down, so can protein expression. This is an attractive technique for scientists who are seeking to better understand the importance of the target gene function by 'knocking down' gene expression in a cell system. | The native RNAi pathway involves a grooming process where long double-stranded RNA (dsRNA) molecules are refined into double stranded RNA of ~20 nucleotides (first step in the figure to the right). However, the ~20 nucleotide double strand RNA (a substrate for Dicer/R2D2) is now a conventional research product known as siRNA. These ~20 nucleotide siRNA inside a cell is catalytic unwound and one of the two strands, known as the guide strand or antisense strand, is then loaded into the RNA-induced silencing complex (RISC). Experimentally, any mRNA that shares 100% sense compliment sequence to the 'loaded' RISC complex, will bind and undergo nuclease-dependent degradation. As mRNA levels go down, so can protein expression. This is an attractive technique for scientists who are seeking to better understand the importance of the target gene function by 'knocking down' gene expression in a cell system. | ||
