PCR-based generation of shRNA libraries from cDNAs.

ABSTRACT:

Background

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.

Results

We report here an improved method for constructing genome-wide shRNA libraries enzymatically. The method

SUBMITTER: Du C 

PROVIDER: S-EPMC1533825 | biostudies-literature | 2006 Jun

REPOSITORIES: biostudies-literature