重组酶
Cre重组酶
DNA
生物
基因
FLP-FRT重组
克隆(编程)
细胞生物学
位点特异性重组
酶
遗传学
重组
计算生物学
生物化学
转基因
遗传重组
计算机科学
转基因小鼠
程序设计语言
作者
Wesleigh F. Edwards,Douglas D. Young,Alexander Deiters
摘要
Cre recombinase catalyzes DNA exchange between two conserved lox recognition sites. The enzyme has extensive biological application, from basic cloning to engineering knock-out and knock-in organisms. Widespread use of Cre is due to its simplicity and effectiveness, but the enzyme and the recombination event remain difficult to control with high precision. To obtain such control we report the installation of a light-responsive o-nitrobenzyl caging group directly in the catalytic site of Cre, inhibiting its activity. Prior to irradiation, caged Cre is completely inactive, as demonstrated both in vitro and in mammalian cell culture. Exposure to non-damaging UVA light removes the caging group and restores recombinase activity. Tight spatio-temporal control over DNA recombination is thereby achieved.
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