Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, USA.
Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, USA.
Curr Opin Struct Biol. 2024 Oct;88:102878. doi: 10.1016/j.sbi.2024.102878. Epub 2024 Jul 18.
Cre recombinase is a phage-derived enzyme that has found utility for precise manipulation of DNA sequences. Cre recognizes and recombines pairs of loxP sequences characterized by an inverted repeat and asymmetric spacer. Cre cleaves and religates its DNA targets such that error-prone repair pathways are not required to generate intact DNA products. Major obstacles to broader applications are lack of knowledge of how Cre recognizes its targets, and how its activity is controlled. The picture emerging from high resolution methods is that the dynamic properties of both the enzyme and its DNA target are important determinants of its activity in both sequence recognition and DNA cleavage. Improved understanding of the role of dynamics in the key steps along the pathway of Cre-loxP recombination should significantly advance our ability to both redirect Cre to new sequences and to control its DNA cleavage activity in the test tube and in cells.
Cre 重组酶是一种源自噬菌体的酶,已被广泛用于精确操作 DNA 序列。Cre 识别并重组由反向重复和不对称间隔区组成的loxP 序列对。Cre 切割并重新连接其 DNA 靶标,因此不需要易错修复途径来生成完整的 DNA 产物。广泛应用的主要障碍是缺乏对 Cre 如何识别其靶标以及其活性如何受到控制的了解。从高分辨率方法得出的图片表明,酶和其 DNA 靶标的动态特性是其在序列识别和 DNA 切割中的活性的重要决定因素。深入了解动力学在 Cre-loxP 重组途径关键步骤中的作用,应能显著提高我们重新定向 Cre 到新序列的能力,并控制其在试管和细胞中的 DNA 切割活性。