Constructing a Far-Red Light-Induced Split-Cre Recombinase System for Controllable Genome Engineering

Cre/loxP-mediated recombination has been used to manipulate genomic DNA in a variety of organisms. However, a large number of complex biomedical studies require the development of more precise inducible Cre/loxP systems that can spatiotemporally control genomic DNA to better dissect specific gene functions. This chapter describes in detail the protocols of constructing a far-red light (FRL)-induced split-Cre recombinase system for controllable genome engineering, including rational design, optimization, and construction of gene modules for an FRL-induced split Cre-loxP system (FISC), cell transfection, and SEAP reporter assay, as well as the characterization of the FISC system and related experimental procedures for FISC-medicated DNA recombination in mice using hydrodynamic injection or adeno-associated virus (AAV) delivery.