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CreLite : An optogenetically controlled Cre/ loxP system using red light
Author(s) -
Yen ShuoTing,
Trimmer Kenneth A.,
AboulFettouh Nader,
Mullen Rachel D.,
Culver James C.,
Dickinson Mary E.,
Behringer Richard R.,
Eisenhoffer George T.
Publication year - 2020
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.232
Subject(s) - biology , zebrafish , cre recombinase , microbiology and biotechnology , green fluorescent protein , gene expression , gene , fate mapping , regulation of gene expression , reporter gene , fusion protein , transgene , embryonic stem cell , genetics , genetically modified mouse , recombinant dna
Background Precise manipulation of gene expression with temporal and spatial control is essential for functional analysis and determining cell lineage relationships in complex biological systems. The cyclic recombinase (Cre)‐ lox P system is commonly used for gene manipulation at desired times and places. However, specificity is dependent on the availability of tissue‐ or cell‐specific regulatory elements used in combination with Cre. Here, we present CreLite, an optogenetically controlled Cre system using red light in developing zebrafish embryos. Results Cre activity is disabled by splitting Cre and fusing with the Arabidopsis thaliana red light‐inducible binding partners, PhyB and PIF6. Upon red light illumination, the PhyB‐CreC and PIF6‐CreN fusion proteins come together in the presence of the cofactor phycocyanobilin (PCB) to restore Cre activity. Red light exposure of zebrafish embryos harboring a Cre‐dependent multicolor fluorescent protein reporter injected with CreLite mRNAs and PCB resulted in Cre activity as measured by the generation of multispectral cell labeling in several different tissues. Conclusions Our data show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different developmental stages, and suggests CreLite may also be useful for temporal and spatial control of gene expression in cell culture, ex vivo organ culture, and other animal models.