PCR-based generation of shRNA libraries from cDNAs

摘要: 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 siRNA libraries emerged as a viable approach. Two different methods have been used generate short hairpin RNA (shRNA) libraries; one is chemically synthesized oligonucleotides, and the other convert complementary DNAs (cDNAs) into shRNA cassettes enzymatically. The high cost chemical synthesis low efficiency enzymatic approach hampered widespread with libraries.