A genetic strategy for stochastic gene activation with regulated sparseness (STARS).
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 sparseness level.
Wang
Sheng-Zhi
SZ
Zilkha Neurogenetic Institute, University of Southern California, Los Angeles, California, United States of America.
Liu
Bao-Hua
BH
Tao
Huizhong W
HW
Xia
Kun
K
Zhang
Li I
LI
D000818:Animals; D002460:Cell Line; D019426:Integrases; D051379:Mice; D011995:Recombination, Genetic; D013269:Stochastic Processes; D015533:Transcriptional Activation; D014162:Transfection / Q000379:methods*
C045073:Cre recombinase; D019426:Integrases