Selection–subtraction approach (SSA): A universal genetic screening technique that enables negative selection

摘要: Screening of expression libraries for bioactive clones that modulate the growth mammalian cells has been limited largely to positive selections incapable revealing suppressive or lethal genetic elements. We have developed a technique, selection–subtraction approach (SSA), allows growth-modulating be isolated based on alterations in their relative abundance growing cell populations transduced with an library. SSA utilizes tagged retroviral bacteriophage λ vectors (retrophages). Nylon prints from retrophage are used determine tags library-transduced identify biological activity individual clones. Applications gene discovery, target and generation mutant proteins demonstrated, by using p53 ataxia telangiectasia mutated (ATM) as models isolate inhibitory proteins, peptides antisense RNAs, temperature-sensitive proteins.