a versatile viral system for expression and depletion of proteins in mammalian cells多才多艺的病毒表达系统和哺乳动物细胞中蛋白质的消耗.pdfVIP

A Versatile Viral System for Expression and Depletion of Proteins in Mammalian Cells 1 1 2,3 1 1 2 Eric Campeau *, Victoria E. Ruhl , Francis Rodier , Corey L. Smith , Brittany L. Rahmberg , Jill O. Fuss , 2,3 2 2 1 Judith Campisi , Paul Yaswen , Priscilla K. Cooper , Paul D. Kaufman 1 Program in Gene Function and Expression, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America, 2 Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, United States of America, 3 Buck Institute for Age Research, Novato, California, United States of America Abstract The ability to express or deplete proteins in living cells is crucial for the study of biological processes. Viral vectors are often useful to deliver DNA constructs to cells that are difficult to transfect by other methods. Lentiviruses have the additional advantage of being able to integrate into the genomes of non-dividing mammalian cells. However, existing viral expression systems generally require different vector backbones for expression of cDNA, small hairpin RNA (shRNA) or microRNA (miRNA) and provide limited drug selection markers. Furthermore, viral backbones are often recombinogenic in bacteria, complicating the generation and maintenance of desired clones. Here, we describe a collection of 59 vectors that comprise an integrated system for constitutive or inducible expression of cDNAs, shRNAs or miRNAs, and use a wide variety of drug selection markers. These vectors are based on the Gateway technology (Invitrogen) whereby the cDNA, shRNA or miRNA of interest is cl