文章基本信息

标题：Improving the Stability of Short Hairpin RNA against Fetal Bovine Serum Using the Third Double-Stranded RNA-Binding Domain from Staufen Protein
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作者：Yasuhisa Kawaguchi ; Kazumi Danjo ; Tomoyuki Okuda 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：2009
卷号：32
期号：2
页码：283-288
DOI：10.1248/bpb.32.283
出版社：The Pharmaceutical Society of Japan
摘要：RNA interference (RNAi) is a technology that specifically inhibits gene expression and is carried out by small 21–27-nucleotide double-stranded small interfering RNA (siRNA) or short hairpin RNA (shRNA). RNAi has become a very promising technology for genetic research, however problems remain with the delivery of siRNA into cells. SiRNA and shRNA are easily degraded by RNases in body fluids and are hardly able to permeate cell membranes because of hydrophilic, polyanionic macromolecules which make their bioavailability very low. We have focused on the third double-stranded RNA-binding domain (dsRBD3) from the Mus musculus Staufen protein in order to develop a non-viral, multifunctional, artificial virus-like delivery system. We constructed a dsRBD3 expression vector and the recombinant dsRBD3 was expressed as a fusion protein with affinity tags. Purified dsRBD3 was mixed with siRNA or shRNA at various ratios and then added to fetal bovine serum (FBS) to evaluate the inhibition of the degradation of double-stranded RNA (dsRNA) by RNase. Unexpectedly, dsRBD3 was not able to protect the siRNA against degradation by FBS, but shRNA was stabilized to some degree by dsRBD3.
关键词：RNA interference;short hairpin RNA;double-stranded RNA-binding domain;Staufen protein;gene delivery;non-viral vector