Large-Scale, High-Throughput Validation of Short Hairpin RNA Sequences for RNA Interference-Reference-Cited by-同舟云学术

Large-Scale, High-Throughput Validation of Short Hairpin RNA Sequences for RNA Interference

Published:2006-02-20 Issue:3 Volume:11 Page:236-246
ISSN:1087-0571
Container-title:Journal of Biomolecular Screening
language:en
Short-container-title:J Biomol Screen

Author:

Lamarcq Laurence H.,Scherer Bradley J.,Phelan Michael L.,Kalnine Nikolai N.,Nguyen Yen H.,Kabakova Tatyana,Chen Xiaoyi,Tan Marcia,Chang Cynthia¹,Berlon Charina,Campos-Gonzalez Roberto,Gao Guo-Jian²,Golz Stefan³,Vysotski Eugene S.⁴,Farmer Andrew A.⁵

Affiliation:

1. Clontech Laboratories, Mountain View, CA

2. BD Biosciences Pharmingen, La Jolla, CA

3. Bayer Healthcare AG, Pharma Research—Molecular Screening Technology, Wuppertal, Germany

4. Russian Academy of Science—Siberian Branch, Institute of Biophysics, Photobiology Lab, Krasnoyarsk, Russia

5. Clontech Laboratories, 1290 Terra Bella Mountain View, CA 94043

Abstract

A method for high-throughput cloning and analysis of short hairpin RNAs (shRNAs) is described. Using this approach, 464 shRNAs against 116 different genes were screened for knockdown efficacy, enabling rapid identification of effective shRNAs against 74 genes. Statistical analysis of the effects of various criteria on the activity of the shRNAs confirmed that some of the rules thought to govern small interfering RNA (siRNA) activity also apply to shRNAs. These include moderate GC content, absence of internal hairpins, and asymmetric thermal stability. However, the authors did not find strong support for positionspecific rules. In addition, analysis of the data suggests that not all genes are equally susceptible to RNAinterference (RNAi).

Publisher

Elsevier BV

Link

http://journals.sagepub.com/doi/pdf/10.1177/1087057105284342

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