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一个小小的新型细菌Eubacterium siraeum的表征和工程 CRISPR-Cas12f系统

Yannan Wang¹, Yujue Wang¹, Na Tang¹

¹School of Physical Science and Technology & State Key Laboratory of Advanced Medical Materials and Devices, ShanghaiTech University, Shanghai 201210, China.

ACS synthetic biology

|June 28, 2024

概括

研究人员从Eubacterium siraeum (EsCas12f1) 设计了一个紧的Cas12f基因组编辑器. 这种新的工具显示了 in vivo 基因编辑和 in vitro 诊断的潜力,扩大了 CRISPR-Cas 工具箱.

科学领域:

分子生物学分子生物学
生物化学生物化学
遗传学遗传学是一个

背景情况:

Cas12f核酶是具有治疗潜力的紧基因组编辑器.
活跃的Cas12f编辑器的可用性是有限的.
需要新的微型Cas12f内核酶.

研究的目的:

描述和设计一种来自Eubacterium siraeum (EsCas12f1) 的新型微型Cas12f内核酶.
为了确定它的原生空间器相邻基因 (PAM) 偏好和生物化学性质.
优化EsCas12f1用于基因组编辑和核酸诊断.

主要方法:

对EsCas12f1内核酶活性的表征.
阐明PAM特异性的解释.
对CRISPR-EsCas12f1系统的引导RNA的合理设计和优化.
在体外核酸诊断的演示.

主要成果:

一个新的微型Cas12f内核酶,EsCas12f1 (433个氨基酸),被确定和描述.
阐明了DNA向和裂变的特定PAM偏好和生化特性.
优化的CRISPR-EsCas12f1系统证明了高效的细菌基因组编辑.
EsCas12f1在体外核酸诊断中显示出有用性.

结论:

EsCas12f1扩展了微型的CRISPR-Cas工具箱.
工程EsCas12f1是一种高效的细菌基因组编辑器.
EsCas12f1对基因组编辑和体外诊断应用都有希望.

关键词:

这就是CRISPR-Cas12f.在gRNA工程方面,基因组编辑基因组编辑在体外诊断 in vitro诊断

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