相关实验视频

Updated: Jul 23, 2025

CRISPR-based Shuttle Cloning: A High-throughput Cloning Method

CRISPR-based Shuttle Cloning: A High-throughput Cloning Method

Published on: June 13, 2025

一个体外CRISPR-Cas12a介导的协议,用于直接克隆大型DNA片段

Xiaoqian Zeng¹, Shuliu Wang¹, Mindong Liang¹

¹State Key Laboratory of Bioreactor Engineering, and School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.

|July 11, 2023

概括

克隆大型生物合成基因集群 (BGCs) 对药物发现至关重要,但对于高GC含量细菌来说很困难. 这项研究介绍了一种体外CRISPR-Cas12a方法,用于高效地直接克隆来自这些具有挑战性的生物体的大型DNA片段.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
基因组学就是基因组学.

背景情况:

发现基于天然产品的新药依赖于克隆大型生物合成基因集群 (BGCs).
克隆大型BGC在高GC含量的微生物中尤其具有挑战性,例如Actinobacteria.

研究的目的:

开发一种高效的体外克隆方法,用于直接克隆大型DNA片段.
从高GC含量的微生物中克隆大型BGC的局限性.

主要方法:

使用了体外CRISPR-Cas12a系统进行向DNA裂变.
详细的协议包括crRNA设计,DNA分离,CRISPR-Cas12a裂变和捕获等离子体的构造.
使用结合,转化和选来成功克隆目标BGCs.

主要成果:

成功演示了一种体外CRISPR-Cas12a介导的协议,用于直接克隆大型DNA片段.
该协议有助于从高GC含量细菌中克隆具有挑战性的大型BGC.

结论:

提出的CRISPR-Cas12a协议为难以在微生物宿主中进行大型BGC克隆提供了一个可行的解决方案.
这种方法可以加快药物开发的新天然产品的发现.

关键词:

生物技术和生物工程克里斯普尔是什么意思?克里斯普尔是什么意思?

更多相关视频

A Universal Protocol for Large-scale gRNA Library Production from any DNA Source

A Universal Protocol for Large-scale gRNA Library Production from any DNA Source

Published on: December 6, 2017

Author Spotlight: Simplifying Genome-Wide Plasmid Library Construction Using CRISPRmass

Author Spotlight: Simplifying Genome-Wide Plasmid Library Construction Using CRISPRmass

Published on: May 17, 2024

相关概念视频

DNA Isolation

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...