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Anal Biochem. 2013 Jul 1;438(1):39-41. doi: 10.1016/j.ab.2013.03.011. Epub 2013 Mar 20.

A single-step mixing cloning method for assembly of lentiviral short hairpin RNA expression vectors for gene silencing.

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  • 1Institute of Biochemistry and Molecular Biology, Hubei University, Wuhan 430062, Hubei Province, People's Republic of China.

Abstract

Lentiviral expression vectors encoding short hairpin RNA (shRNA) are widely used for RNAi-based gene silencing in mammalian cells. However, current methods for the construction of shRNA expression vectors require multiple steps, which are expensive, time-consuming, and error-prone. Here, we developed a single-step mixing cloning method for the generation of lentiviral shRNA expression vectors. With this method, a pair of short oligonucleotides (∼50 nt) is required and a lentiviral shRNA vector can be constructed with only one step. This method has been used to construct 30 lentiviral shRNA expression vectors successfully.

Copyright © 2013 Elsevier Inc. All rights reserved.

PMID:
23524019
[PubMed - indexed for MEDLINE]
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