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exoCasMINI:一种T5外核酶融合的CRISPR-Cas12f系统,具有增强的基因编辑效率

Kaitong Liao¹, Kechen Chen¹, Shufeng Ma^1,2

¹Cancer Research Institute, School of Basic Medical Sciences, State Key Laboratory of Multi-organ Injury Prevention and Treatment, Guangdong Province Key Laboratory of Immune Regulation and Immunotherapy, Southern Medical University, Guangzhou 510515, China.

|August 12, 2025

概括

工程 Cas12f 系统 (exoCasMINI) 提高了基因组编辑的效率和特异性,克服了较大的 CRISPR 系统的局限性. 这些新型工具对哺乳动物细胞的基因治疗应用有前途.

科学领域:

分子生物学分子生物学
基因编辑技术的技术
生物技术是生物技术.

背景情况:

克里斯普尔-Cas9和Cas12a是标准的基因组编辑器,但由于它们的尺寸,它们面临着交付挑战.
紧的Cas12f蛋白质提供更好的传递,但编辑活动有限.

研究的目的:

设计增强的Cas12f系统,以提高基因组编辑的活性和特异性.
评估工程系统在哺乳动物细胞和体内生物中的效率和特异性.

主要方法:

将T5外核酶与CasMINI的融合形成exoCasMINI.
与SpCas9,LbCas12a和CasMINI相比,评估编辑效率和特异性.
通过整合KrasG12D和破坏瘤抑制剂来诱导瘤发生的exoCasMINI体内评估.

主要成果:

exoCasMINI显示的编辑效率比CasMINI高1.1-21.1倍,与SpCas9和LbCas12a活动相匹配.
与SpCas9.9相比,工程系统诱导了更长的删除,并显示出更高的特异性.
在小鼠肝脏模型中,exoCasMINI在诱导瘤发生方面比CasMINI更有效.
改造后的exoRhCas12f1也显示出显著增强的活性 (1.2-3.6倍).

结论:

工程化 exoCasMINI 和 exoRhCas12f1 是高效和特定的基因组编辑工具.
这些紧的系统克服了交付限制,并比现有的CRISPR技术提供了优势.
增强的Cas12f系统对未来的基因治疗应用具有重大潜力.

关键词:

生物化学的应用生物化学方法生物化学方法酶学是一种酶学.基因学的技术在遗传学.

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