@article{100581f10e18432596a93428af8a34a1,
title = "CreLite: An optogenetically controlled Cre/loxP system using red light",
abstract = "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)-loxP 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.",
keywords = "Cre/loxP, gene expression, optogenetics, zebrafish",
author = "Yen, {Shuo Ting} and Trimmer, {Kenneth A.} and Nader Aboul-Fettouh and Mullen, {Rachel D.} and Culver, {James C.} and Dickinson, {Mary E.} and Behringer, {Richard R.} and Eisenhoffer, {George T.}",
note = "Funding Information: The authors thank Dr. David Hawk from the Proteomics and Metabolomics Core of MD Anderson Cancer Center for helping the HPLC purification of Phycocyanobilin. The authors also thank Dr. Adriana Paulucci in the Microscope core of the Genetics Department at the MD Anderson Cancer Center and Jordan Pietz with MD Anderson Creative Communications group for assistance with the biomedical illustrations. This work was supported by National Institutes of Health (NIH) grant OD19764 and the Ben F. Love Endowment to R. R. B.; by the NIH training grant, T32HL007676, to J. C. C.; and by the Cancer Prevention Institute of Texas, RR140077, National Institutes of General Medical Sciences, R01GM124043, and the Mark and Linda Quick Basic Science Award to G. T. E. Funding Information: The authors thank Dr. David Hawk from the Proteomics and Metabolomics Core of MD Anderson Cancer Center for helping the HPLC purification of Phycocyanobilin. The authors also thank Dr. Adriana Paulucci in the Microscope core of the Genetics Department at the MD Anderson Cancer Center and Jordan Pietz with MD Anderson Creative Communications group for assistance with the biomedical illustrations. This work was supported by National Institutes of Health (NIH) grant OD19764 and the Ben F. Love Endowment to R. R. B.; by the NIH training grant, T32HL007676, to J. C. C.; and by the Cancer Prevention Institute of Texas, RR140077, National Institutes of General Medical Sciences, R01GM124043, and the Mark and Linda Quick Basic Science Award to G. T. E. Funding Information: Cancer Prevention and Research Institute of Texas, Grant/Award Number: RR140077; National Heart, Lung, and Blood Institute, Grant/Award Number: T32HL007676; National Institute of General Medical Sciences, Grant/Award Number: R01GM124043; NIH Office of the Director, Grant/Award Number: OD19764; Mark and Linda Quick Basic Science Award; Ben F. Love Endowment Funding information Publisher Copyright: {\textcopyright} 2020 Wiley Periodicals LLC",
year = "2020",
month = nov,
day = "1",
doi = "10.1002/dvdy.232",
language = "English (US)",
volume = "249",
pages = "1394--1403",
journal = "Developmental Dynamics",
issn = "1058-8388",
publisher = "Wiley-Liss Inc.",
number = "11",
}