Short Interfering RNA

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Background Information

RNA inference (RNAi) was first discovered in Cenorhabditis elegans nearly a decade ago and have been revolutionizing gene-function analysis ever since. This discovery began the process by which scientists work with a known gene sequence and attempt to define its biological function by disrupting its activity in vivo. It involves the introduction of homologous double stranded RNA (dsRNA) to specifically target a gene's product and to disrupt the function of that gene in vivo. The resulting effects on an animal are referred to as a phenocopy due to the fact that it copies the phenotype of a loss-of-function mutation of that gene, but is not really inherited like how a true gene deletion would be.

Despite the variety of technologies, this was not possible before due to the lack of genome information. The advances in gene sequencing throughout the past couple of years have allowed various usages of available technology to go after any gene and try to identify the gene, as well as to understand its specific function. Once the sequence is known, an open reading frame can be obtained to find a specific coding region of a gene by looking for the universal start codon, AUG, and the universal stopping codon, UAA, UUA, or UAG. From there, RNAi can be introduced into cells of certain organisms, with dsRNA as the interfering agent that can result in the destruction and degradation of a sequence specific messenger RNA (mRNA) that mimics a loss-of-function phenotype. In other words, RNAi is a very efficient and highly specific way in which to inactivate gene function and can serve as an alternative to gene knockouts, or the conventional antisense technology.

The mechanism of how dsRNA results in...

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...of a marker gene and a hairpin expression cassette in a viral vector would allow single-cell or mosaic analysis of gene function.

• It may be possible in the future to apply this process of hairpin siRNA synthesis to studies of neurogenesis and differentiation in mammals.

The information contained in this website comes from (or was sought out because of) the paper entitled "RNA interference by expression of short-interfering RNAs and hairpin RNAs in mammalian cells" written by Dr. Jenn-Yah Yu, Dr. Stacy L. DeRuiter, and Dr. David L. Turner at the University of Michigan's Department of Biological Chemistry.

Citations:

DeRuiter, Stacy. Yu, Jenn-Yah. Turner, David L. "RNA interference by expression of short-interfering RNAs and hairpin RNAs in mammalian cells." Proceedings of the National Academy of Sciences, USA. Vol. 99, Issue 9, 6047-6052, April 30, 2002