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开发和实施一种基于CRISPR的类型I-C可编程抑制系统,用于Neisseria gonorrhoeae

Wendy E Geslewitz¹, Amaris Cardenas², Xufei Zhou³

¹Department of Microbiology and Immunology, Northwestern University, Chicago, Illinois, USA.

|December 21, 2023

概括

我们使用Neisseria lactamica I-C型CRISPR-Cas系统开发了针对尼塞利亚淋病的CRISPR干扰 (CRISPRi) 系统. 这种工具允许基因抑制,有助于研究必要的基因和开发新的淋病治疗方法.

科学领域:

微生物学微生物学
分子生物学分子生物学
遗传学遗传学是一种遗传学.

背景情况:

集群定期间隔的短平行体重复 (CRISPR) - - 卡斯系统是原生生物的自适应性免疫机制.
尼塞利亚淋病 (Gc) 缺乏内生CRISPR系统,阻碍了基因操纵.
乳糖菌 (Neisseria lactamica) 具有一个功能性的I-C型CRISPR-Cas系统.

研究的目的:

开发一种可编程和可诱导的CRISPR干扰 (CRISPRi) 平台,用于Gc.中的基因抑制.
建立一种工具,用于在GC中询问基本和非基本基因.
为了促进研究GC生理学,病变发生和抗微生物开发.

主要方法:

来自N. lactamica型I-C CRISPR的异烯β-d-1-thiogalactopyranoside (IPTG) -可诱导的CRISPRi系统,缺乏Cas3核酶.
针对特定的基因,包括opaD基因和整个opa基因家族,用于转录抑制.
评估了基因和蛋白质表达,细菌表型,以及在IPTG去除后抑制的可逆性.

主要成果:

证明成功降低OPAD基因和蛋白质的表达,影响细菌的功能.
通过使用五位空间CRISPR阵列,有效地击败了所有11个OPA基因.
确认可逆的基因抑制和基本基因表达的有条件减少.

结论:

开发的I-C型CRISPRi系统为Gc.中的特定位置的转录抑制提供了一个强大的工具.
这种系统使得基本基因的功能特征能够被确定,而这些基因在其他情况下很难被研究.
克里斯皮尔平台具有很大的潜力,可以推进GC研究,并帮助开发新疗法.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?尼塞里亚淋病菌 (Neisseria gonorrhoeae) 是一种传染病.大大大大大大大大大大大大大大大大大大大大大大大大大基本的基因基本的基因中性粒细胞中性粒细胞.在这里,我们可以看到PiliPiliPili.转录规则转录规则转录规则

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