克里斯皮尔i-TnSeq:一种全基因组高通量工具,用于绘制细菌基本与非基本遗传相互作用的地图
bioRxiv : the preprint server for biology
|July 3, 2023
概括
我们开发了CRISPRi-TnSeq,这是一个新的工具,用于绘制细菌中必需基因和非必需基因之间的遗传相互作用. 这种方法揭示了功能性基因关系,有助于理解基因功能和识别潜在的药物标.
科学领域:
- 微生物学 微生物学
- 系统生物学 系统生物学
- 基因组学就是基因组学.
背景情况:
- 基因相互作用网络对于理解基因功能,药物标和途径完成至关重要.
- 现有的工具缺乏在各种细菌菌株和物种中绘制遗传相互作用的能力.
研究的目的:
- 开发和验证CRISPRi-TnSeq,这是一个全基因组工具,用于绘制基本基因和非基本基因之间的遗传相互作用.
- 在*Streptococcus pneumoniae*中构建和分析基本-非基本的遗传相互作用网络.
- 发现新的基因功能,途径依赖性和潜在的治疗点.
主要方法:
- CRISPRi-TnSeq结合了CRISPR干扰 (CRISPRi) 进行基本基因淘汰,并将转子子序列 (Tn-Seq) 进行非基本基因淘汰.
- 在 *Streptococcus pneumoniae* 中,为13个重要的基因生成了CRISPRi菌株.
- 对转子子突变库进行了选,以确定大约24,000个基因-基因对,揭示了1,334个遗传相互作用.
主要成果:
- 在基本基因和非基本基因之间确定了1334个遗传相互作用,包括754个负面相互作用和580个正面相互作用.
- 发现了17个类基因,其中一些作为缓冲干扰的遗传电容器.
- 详细的特定途径相互作用涉及细胞壁合成,分裂和c-di-AMP信号传递.
结论:
- CRISPRi-TnSeq有效地揭示了基因相互作用,揭示了途径依赖性,并提供了对基因功能的洞察.
- 该方法依赖于广泛使用的技术,使其适应于在各种微生物物种中构建基因相互作用网络.
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