Journal
ANALYTICAL BIOCHEMISTRY
Volume 349, Issue 1, Pages 41-48Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2005.11.005
Keywords
RNA interference; Alu; SINE; shRNA; SeshRNA; retroelement
Funding
- NIGMS NIH HHS [R01GM45668] Funding Source: Medline
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RNA interference has become a powerful tool for specific inhibition of gene expression in mammalian cells. Expression constructs allow for the long-term delivery of short interfering RNAs, usually through the expression of Pol III-transcribed hairpins. In some instances, these expression systems have been shown to have side effects, including induction of the interferon response and cytotoxicity. Here we demonstrate that HI-expressed hairpins, as well as the cloning vector, reduce the plating efficiency of HeLa cells. This toxicity is abrogated by coexpression of the hairpin in the same transcript as a human Alu repetitive element. These Alu-linked hairpins retain the ability to knock down expression of target mRNAs. This modification, which we term SINE (short interspersed repetitive element)enhanced short hairpin RNA, provides an alternative expression system for hairpins with reduced side effects. (c) 2005 Elsevier Inc. All rights reserved.
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