克里斯普尔混合:一种克里斯普尔介导的细胞内定向进化平台,用于RNA适应体
bioRxiv : the preprint server for biology
|September 11, 2023
概括
研究人员开发了一种新的CRISPR系统,以进化RNA吸收体,以精确调节基因. 这种方法使哺乳动物细胞中的多个基因能够同时激活和抑制,使用新型的aptamer-RNA结合蛋白对.
科学领域:
- 分子生物学分子生物学
- 合成生物学 合成生物学
- 基因规则 基因规则
背景情况:
- 克里斯普技术为基因编辑和表达调节提供了强大的工具.
- 将RNA吸附体与CRISPR的单导向RNA (sgRNA) 融合,可以招募RNA结合蛋白 (RBPs) 来准基因组部位.
- 多重复合基因调节可以通过表达具有多种RNA体的sgRNA来实现.
研究的目的:
- 开发一种细胞内定向进化平台,用于向细胞内RBP的RNA吸收体.
- 为了识别与现有的aptamer-RBP对正交的新型RNA体.
- 为了证明这些直角的阿普坦-RBP对在多重化CRISPR基因调节中的应用.
主要方法:
- 优化一种与光激活细胞分类 (FACS) 结合的细菌CRISPR混合系统.
- 定向进化选择高亲和度RNA吸收体对抗细胞内RBP.
- 在哺乳动物细胞的多重CRISPR系统中应用选择的正交的阿普坦-RBP对.
主要成果:
- 成功建立了RNA体的细胞内定向进化平台.
- 鉴定具有高亲和度和对特定RBP的正交度的新型RNA吸收体.
- 使用多重化CRISPR与正交的阿普坦-RBP对同时实现内源基因的转录激活和抑制的演示.
结论:
- 开发的平台可以发现用于基因调节的功能性RNA体.
- 多重复合的CRISPR应用利用直角的阿帕特马-RBP对允许同时转录激活和抑制哺乳动物细胞内源基因.
- 这项工作推进了基于CRISPR的工具对复杂基因操纵策略的潜力.
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