Protocol for NT-CRISPR: A Method for Efficient Genome Engineering in Vibrio natriegens.

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/978-1-0716-4220-7_20

Daniel Stukenberg, Josef Hoff, Anna Faber, Anke Becker

引用次数: 0

Abstract

Vibrio natriegens is a gram-negative bacterium, which has received increasing attention due to its very fast growth with a doubling time of under 10 min under optimal conditions. To enable a wide range of projects spanning from basic research to biotechnological applications, we developed NT-CRISPR as a new method for genome engineering. This book chapter provides a step-by-step protocol for the use of this previously published tool. NT-CRISPR combines natural transformation with counterselection through CRISPR-Cas9. Thereby, genomic regions can be deleted, foreign sequences can be integrated, and point mutations can be introduced. Furthermore, up to three simultaneous modifications are possible.

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NT-CRISPR：一种高效的纳氏弧菌基因组工程方法。

纳氏弧菌（Vibrio natriegens）是一种革兰氏阴性细菌，由于其生长速度极快，在最佳条件下翻倍时间不到 10 分钟，因此受到越来越多的关注。为了实现从基础研究到生物技术应用的广泛项目，我们开发了 NT-CRISPR，作为基因组工程的一种新方法。本书的这一章将逐步介绍如何使用这一之前已发表的工具。NT-CRISPR 将自然转化与 CRISPR-Cas9 的反选择相结合。因此，可以删除基因组区域、整合外来序列并引入点突变。此外，最多可同时进行三种改造。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.