CRISPRe: An innate transcriptional enhancer for endogenous genes in CRISPR-Cas immunity,iScience

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CRISPRe: An innate transcriptional enhancer for endogenous genes in CRISPR-Cas immunity
iScience ( IF 4.6 ) Pub Date : 2023-09-01 , DOI: 10.1016/j.isci.2023.107814
Dan Li _{1,

2} , Yihong Chen ₁ , Fei Huang ₁ , Jianmei Wang ₁ , Xufeng Li ₁ , Yi Yang ₁

Affiliation

CRISPR-Cas system has been repurposed to the promising strategy of CRISPR-based transcriptional interference/activation (CRISPRi/CRISPRa) without eliciting DNA breaks that enables Cas complex a block for transcription initiation or elongation, which greatly expands its application fields and values. However, loss of Cas nuclease ability, especially the endogenous nuclease, may affect genome stability seriously. Here, we found a transcriptional enhancer for genes (CRISPRe) in type I-C system of industrial strain by maintaining the natural activity of Cas3 nuclease and introducing the specific motifs that do not trigger immunity. CRISPRe greatly improved the expression of heterologous and endogenous genes and the biosynthesis of products by facilitating transcriptional elongation. Besides, the mechanism for pyrroloquinoline quinone (PQQ) biosynthesis regulated by coupling transcriptional-translational elongation in operon was elucidated. Hence, we enrich the toolbox for CRISPR-Cas system and provide a new framework for gene regulation at transcription.

中文翻译：

CRISPRe：CRISPR-Cas免疫中内源基因的先天转录增强子

CRISPR-Cas系统已被重新用于基于CRISPR的转录干扰/激活（CRISPRi/CRISPRa）的有前途的策略，而不引起DNA断裂，使Cas能够复合转录起始或延伸的块，这极大地扩展了其应用领域和价值。然而，Cas核酸酶能力的丧失，尤其是内源性核酸酶的丧失，可能会严重影响基因组的稳定性。在这里，我们通过保持Cas3核酸酶的天然活性并引入不触发免疫的特定基序，在工业菌株的I-C型系统中发现了基因转录增强子（CRISPRe）。 CRISPRe通过促进转录延伸极大地改善了异源和内源基因的表达以及产物的生物合成。此外，还阐明了操纵子中转录-翻译延伸耦合调控吡咯喹啉醌（PQQ）生物合成的机制。因此，我们丰富了CRISPR-Cas系统的工具箱，并为转录时的基因调控提供了新的框架。

更新日期：2023-09-01

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