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Updated: Sep 12, 2025

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保护性抗原介导的反CRISPR蛋白的输送用于精确的基因组编辑

Axel O Vera^1,2, Nicholas L Truex¹, Vedagopuram Sreekanth^2,3,4

¹Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139.

Proceedings of the National Academy of Sciences of the United States of America

|August 4, 2025

概括

一种新的蛋白质输送系统,LF_N-Acr/PA,使抗CRISPR蛋白 (Acrs) 能够快速进入细胞,以精确控制Cas9活性. 这提高了CRISPR基因组编辑的安全性和有效性,通过尽量减少不必要的遗传修改.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
基因编辑基因编辑

背景情况:

克里斯普尔-Cas9技术提供了强大的基因组编辑,但需要精确的剂量控制,以防止异常效应,马赛克主义和基因毒性.
第二种类型的抗CRISPR蛋白 (Acrs) 可以抑制Cas9的活性,但由于膜不透性和当前的方法 (如病毒载体或电穿孔) 的局限性,它们的传递到细胞中具有挑战性.

研究的目的:

为抗CRISPR蛋白 (Acrs) 开发一个快速有效的细胞透的输送平台,以提高CRISPR-Cas9基因组编辑的安全性和精度.
创建一个基于蛋白质的系统,克服现有的Acr传递方法的局限性,用于治疗应用.

主要方法:

开发了LF_N-Acr/PA,一种由炭毒素衍生的两组蛋白系统,利用细胞进入的保护性抗原 (PA) 和工程Acr (LF_N-Acr).
证明了LF_N-Acr/PA在几分钟内将LF_N-Acr输入各种人体细胞类型的能力,包括不朽细胞系,胚胎干细胞和3D培养物.
量化了在低LF_N-Acr度下抑制Cas9介导的编辑 (淘汰赛,淘汰赛,转录激活,基编辑).

主要成果:

在低至2.5 pM的度下,LF_N-Acr/PA有效地将LF_N-Acr输送到人体细胞中.
达到高达95%的Cas9-介导基因组编辑功能的抑制.
证明LF_N-Acr的定时递送减少了非目标基编辑,并将Cas9的特异性提高了41%.

结论:

到目前为止,LF_N-Acr/PA代表了迄今为止最强大的已知细胞透CRISPR-Cas抑制系统.
该平台显著提高了CRISPR-Cas9基因组编辑的精度和安全性,扩大了其治疗潜力.
快速交付机制在现有方法上为实时控制Cas9活动提供了显著的优势.

关键词:

克里斯普尔-Cas9是什么意思炭病保护性抗原炭病保护性抗原这是一个反-CRISPR.精确的基因组编辑.提供蛋白质的蛋白质输送.

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