Endoribonuclease-prepared siRNA
esiRNA or Endoribonuclease-prepared siRNAs are a mixture of siRNA oligos resulting from cleavage of long double-stranded RNA (dsRNA) with an endoribonuclease such as Escherichia coli RNase III or Dicer.[1][2]
An alternative concept to the usage of chemically synthesized siRNA for RNA Interference (RNAi) is the enzymatic digestion of long double stranded RNAs in vitro. In this case a cDNA template is amplified by PCR and tagged with two bacteriophage-promoter sequences. RNA polymerase is then used to generate long double stranded RNA that is homologous to the target-gene cDNA. This RNA is subsequently digested with RNase III from Escherichia coli to generate short overlapping fragments of siRNAs with a length between 18-25 base pairs. This complex mixture of short double stranded RNAs is similar to the mixture generated by Dicer cleavage in vivo and is therefore called endoribonuclease-prepared siRNA or short esiRNA. esiRNA are a heterogeneous mixture of siRNAs that all target the same mRNA sequence. These multiple silencing triggers lead to highly specific and effective gene silencing.[2]
References
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- Yang, D.; et al. (2002). "Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells". Proc. Natl. Acad. Sci. U.S.A. 99 (15): 9942–7. doi:10.1073/pnas.152327299. PMC 126604. PMID 12096193.