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

Meiyan Wang; Jiali Wu; Haifeng Ye

doi:10.1201/b22823-10

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

Meiyan Wang
, Jiali Wu
, Haifeng Ye

School of Life Sciences

East China Normal University

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

Abstract

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.

Original language	English
Title of host publication	Opsin-free Optogenetics
Subtitle of host publication	Technology and Applications
Publisher	CRC Press
Pages	135-149
Number of pages	15
ISBN (Electronic)	9781000787542
ISBN (Print)	9781032249223
DOIs	https://doi.org/10.1201/b22823-10
State	Published - 1 Jan 2022

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title = "Constructing a Far-Red Light-Induced Split-Cre Recombinase System for Controllable Genome Engineering",

abstract = "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.",

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AB - 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.

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ER -

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

Abstract

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