Cas Protein - Cas9蛋白作用机制详解 - 知乎 - 知乎专栏
2021年6月17日 · 博德研究所的研究人员对Cas9进行突变使RuvC和HNH两个催化结构域中的一个缺失核酸酶活性形成Cas9n，Cas9n与DNA双链作用时仅产生单链切口。 应用 …
Structure and Dynamics of Cas9 HNH Domain Catalytic State
2017年12月8日 · Using two distinct molecular dynamics simulation techniques, we have obtained a cross-validated catalytically active state of Cas9 HNH domain primed for cutting the target …
Structural insights into DNA cleavage activation of CRISPR-Cas9 …
2017年11月9日 · Here, we report a 5.2 Å cryo-EM structure of Cas9 in complex with sgRNA and target DNA. This structure reveals a conformational state of Cas9 in which the HNH domain is …
Substrate-independent activation pathways of the CRISPR-Cas9 HNH ...
2023年12月19日 · The HNH nuclease of Cas9 is a self-contained catalytic domain for cleavage of target DNA (tDNA) that is basepaired with guide RNA (gRNA) in a heteroduplex (1, 2). In …
Structural determinants of DNA cleavage by a CRISPR HNH …
By contrast, in Cas9, HNH contacts the REC lobe and is inserted in the RuvC domain with two linkers that are close to the DNA unwinding point. These differences in HNH position and …
Exploring alternative catalytic mechanisms of Cas9 HNH domain
Cas9 has two nuclease domains: HNH domain and RuvC domain that are designed to cleave the target DNA (tDNA) strand and the non-target DNA (ntDNA) strand respectively. The HNH …
Coupled catalytic states and the role of metal coordination in Cas9 ...
2023年10月2日 · The HNH domain of Cas9 has a typical ββα-metal fold and contains the conserved Asn (on α) as well as Asp and His (on β1) catalytic residues 13.
Replacing the Sp Cas9 HNH domain by deaminases generates …
2021年12月3日 · In this work, we demonstrate that replacing the HNH domain of SpCas9 with a deaminase domain shifts the editing window of BEs PAM proximally (from positions 4–8 to …
Structural and functional insights into the bona fide catalytic state ...
2019年7月30日 · Our study contributes critical information to defining the catalytic conformation of the HNH domain and advances the knowledge about the conformational activation underlying …
Structural and dynamic insights into the HNH nuclease of divergent Cas9 ...
CRISPR-Cas9 is a widely used biochemical tool with applications in molecular biology and precision medicine. The RNA-guided Cas9 protein uses its HNH endonuclease domain to …