Biochemical reconstitution of a type I-B CRISPR-associated transposon.

4区生物学 Q3 Biochemistry, Genetics and Molecular Biology

Methods in enzymology Pub Date : 2025-01-01 Epub Date: 2025-02-26 DOI:10.1016/bs.mie.2025.01.042

Shukun Wang, Leifu Chang

引用次数: 0

Abstract

CRISPR-associated transposons (CASTs) are potential gene editing tools because of their RNA-guided DNA insertion activity. It is essential to understand the mechanisms underlying the transposition for the application of CASTs. Here, we provide protocols for the biochemical reconstitution of a type I-B CAST for RNA-guided transposition. The procedures may be applicable to other types of CASTs and facilitate the mechanism studies of various CASTs.

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CRISPR相关转座子（CAST）具有RNA引导的DNA插入活性，是一种潜在的基因编辑工具。了解转座子的内在机制对于 CASTs 的应用至关重要。在这里，我们提供了用于 RNA 引导转座的 I-B 型 CAST 的生化重组方案。这些步骤可能适用于其他类型的 CAST，并有助于各种 CAST 的机制研究。

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 enzymology 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

308

审稿时长

3-6 weeks

期刊介绍： The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.