A chemical-induced reversible gene knockout system for studying gene function in situ [RNA-seq]. A chemical-induced reversible gene knockout system for studying gene function in situ [RNA-seq]

We developed a conditional conversion allele system termed CoCo. Gene knockout is achieved through Cre-mediated transcription termination, whereas gene restoration is achieved by Flp-mediated deletion of termination signal. To tightly control the temporal expression of the recombinases, Tet-On inducible system is used to drive the expression of Cre recombinase and tamoxifen-inducible FlpO-ER fusion and therefore a dual drug-inducible conditional conversion system (DDCoCo) is developed. This system provides an efficient, rapid and flexible gene switch for studying gene function. Overall design: 9 samples for High-throughput sequencing, including CC_Cas9_Cre-1, CC_Cas9_Cre-2, CC_Cas9_Cre-3, CC_Cas9_Flpo-1, CC_Cas9_Flpo-2, CC_Cas9_Flpo-3, CC_Cas9-1, CC_Cas9-2, CC_Cas9-3