Short Hairpin RNAs for Strand-Specific Small Interfering RNA Production

Peike Sheng; Krystal A Flood; Mingyi Xie

doi:10.3389/fbioe.2020.00940

Short Hairpin RNAs for Strand-Specific Small Interfering RNA Production

Front Bioeng Biotechnol. 2020 Aug 7:8:940. doi: 10.3389/fbioe.2020.00940. eCollection 2020.

Authors

Peike Sheng^{1

2}, Krystal A Flood^{1

2}, Mingyi Xie^{1

2

3}

Affiliations

¹ Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL, United States.
² UF Health Cancer Center, University of Florida, Gainesville, FL, United States.
³ UF Genetics Institute, University of Florida, Gainesville, FL, United States.

Abstract

RNA interference (RNAi) is an effective mechanism for inhibiting gene expression at the post-transcriptional level. Expression of a messenger RNA (mRNA) can be inhibited by a ∼22-nucleotide (nt) small interfering (si)RNA with the corresponding reverse complementary sequence. Typically, a duplex of siRNA, composed of the desired siRNA and a passenger strand, is processed from a short hairpin RNA (shRNA) precursor by Dicer. Subsequently, one strand of the siRNA duplex is associated with Argonaute (Ago) protein for RNAi. Although RNAi is widely used, the off-target effect induced by the passenger strand remains a potential problem. Here, based on current understanding of endogenous precursor microRNA (pre-miRNA) hairpins, called Ago-shRNA and m⁷G-capped pre-miRNA, we discuss the principles of shRNA designs that produce a single siRNA from one strand of the hairpin.

Keywords: Argonaute; Dicer; RNA interference; m7G-capped pre-miRNA; microRNA; short hairpin RNA; transcription start site miRNA.

Abstract

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