内切酶
生物
基因组
基因
DNA
计算生物学
遗传学
归巢(生物学)
核酸内切酶
系统发育树
生态学
作者
Ryo Takeuchi,Abigail R. Lambert,Amanda Nga-Sze Mak,Kyle Jacoby,Russell J. Dickson,Gregory B. Gloor,Andrew M. Scharenberg,David R. Edgell,Barry Stoddard
标识
DOI:10.1073/pnas.1107719108
摘要
Homing endonucleases mobilize their own genes by generating double-strand breaks at individual target sites within potential host DNA. Because of their high specificity, these proteins are used for “genome editing” in higher eukaryotes. However, alteration of homing endonuclease specificity is quite challenging. Here we describe the identification and phylogenetic analysis of over 200 naturally occurring LAGLIDADG homing endonucleases (LHEs). Biochemical and structural characterization of endonucleases from one clade within the phylogenetic tree demonstrates strong conservation of protein structure contrasted against highly diverged DNA target sites and indicates that a significant fraction of these proteins are sufficiently stable and active to serve as engineering scaffolds. This information was exploited to create a targeting enzyme to disrupt the endogenous monoamine oxidase B gene in human cells. The ubiquitous presence and diversity of LHEs described in this study may facilitate the creation of many tailored nucleases for genome editing.
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