Method for PCR-based generation of shRNA libraries from cDNAs

The use of small interfering RNAs (siRNAs) to silence target gene expression has greatly facilitated mammalian genetic analysis by generating loss-of-function mutants. In recent years, high-throughput, genome-wide screening of siRNA libraries has emerged as a viable approach. Two different methods have been used to generate short hairpin RNA (shRNA) libraries; one is to use chemically synthesized oligonucleotides, and the other is to convert complementary DNAs (cDNAs) into shRNA cassettes enzymatically. The high cost of chemical synthesis and the low efficiency of the enzymatic approach have hampered the widespread use of screening with shRNA libraries.

Reference:

PCR-based generation of shRNA libraries from cDNAs
Cheng Du,Baosheng Ge,Zhongfeng Liu, Kai Fu, Wing C Chan, and Timothy W McKeithan
BMC Biotechnol. 2006; 6: 28

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Last update 12-Dec-2006, Rating n/a of 0 votes.