a genetic strategy for stochastic gene activation with regulated sparseness (stars)遗传策略随机基因激活和调节稀疏(星星).pdfVIP

A Genetic Strategy for Stochastic Gene Activation with Regulated Sparseness (STARS) 1,2 1,2 1,3 4 1,2 Sheng-zhi Wang , Bao-hua Liu , Huizhong W. Tao , Kun Xia , Li I. Zhang * 1 Zilkha Neurogenetic Institute, University of Southern California, Los Angeles, California, United States of America, 2 Department of Physiology Biophysics, Keck School of Medicine, University of Southern California, Los Angeles, California, United States of America, 3 Department of Cell and Neurobiology, Keck School of Medicine, University of Southern California, Los Angeles, California, United States of America, 4 The State Key Laboratory of Medical Genetics, Central South University, Changsha, Hunan, People’s Republic of China Abstract It remains a challenge to establish a straightforward genetic approach for controlling the probability of gene activation or knockout at a desired level. Here, we developed a method termed STARS: stochastic gene activation with genetically regulated sparseness. The stochastic expression was achieved by two cross-linked, mutually-exclusive Cre-mediated recombinations. The stochastic level was further controlled by regulating Cre/lox reaction kinetics through varying the intrachromosomal distance between the lox sites mediating one of the recombinations. In mammalian cell lines stably transfected with a single copy of different STARS transgenes, the activation/knockout of reporter genes was specifically controlled to occur in from 5% to 50% of the cell population. STARS can potentially provide a convenient way for genetic labeling as well as gene expression/knockout in a population of cells with a desired sp