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通过CE-SELEXEX对SpyCas9的aptamer抑制剂的选择

Ge Yang¹, Huimin Wang², Guangyu Jiang³

¹Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, 5 South Zhongguancun Street, Beijing, 100081, China; CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Antimicrobial Agents, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China.

|March 13, 2024

概括

研究人员开发了新型的体来抑制CRISPR-Cas9系统,提供了一个精确可控的基因编辑工具. 这些胺酶抑制剂克服了现有方法对增强CRISPR-Cas9活动控制的局限性.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
基因编辑技术的技术

背景情况:

CRISPR/Cas9是一种广泛用于基因编辑的细菌免疫系统.
现有的CRISPR-Cas9抑制剂 (多,小分子) 有一些局限性,例如难以采购,可用性有限或细胞毒性.
对于CRISPR-Cas9系统,需要新的,有效的和可控制的抑制剂.

研究的目的:

发现和描述针对SpyCas9蛋白质的新型胺酶抑制剂.
开发一种简单,精确和通用的策略来控制CRISPR-Cas9活动.
探索体作为研究CRISPR/Cas9功能的工具的潜力.

主要方法:

结合的基于乳液的电泳分离 (CE-SELEX) 采用一次性压力可控制的选择策略.
实验室酶定量测试以评估已识别的aptamer的抑制活性.
生物信息学预测的aptamer-目标相互作用.

主要成果:

针对SpyCas9成功识别了aptamers,其中Apt2表现出高亲和力 (纳米级).
在实验室中,Apt2证明有效抑制了SpyCas9的酶活性.
预计Apt2将与HNH和RuvC域结合,从而竞争性地抑制基质DNA结合.

结论:

亚胺,特别是Apt2,代表了对SpyCas9.9的一种有前途的寡核酸抑制剂类.
这些体为CRISPR-Cas9活动的普遍,简单和精确控制提供了潜在的策略.
开发的体可以作为研究CRISPR/Cas9机制和功能的宝贵工具.

关键词:

亚普特美尔 (Aptamer) 是一种药物.在CE-SELEXEX中使用.这就是CRISPR/Cas9的作用.抑制剂是一种抑制剂.

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CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced Short...