Simple and Rapid Assembly of TALE Modules Based on the Degeneracy of the Codons and Trimer Repeats

Transcription activator-like effectors (TALEs) have been effectively used for targeted genome editing, transcriptional regulation, epigenetic modification, and locus-specific DNA imaging. However, with the advent of the clustered regularly interspaced short palindromic repeat/Cas9 system, an easy-to...

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Autores principales:	Lingyin Cheng, Xiaoqing Zhou, Yuling Zheng, Chengcheng Tang, Yu Liu, Shuwen Zheng, Yang Liu, Jizeng Zhou, Chuan Li, Min Chen, Liangxue Lai, Qingjian Zou
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	transcription activator-like effectors degenerated codon trimer repeats Gibson assembly Genetics QH426-470
Acceso en línea:	https://doaj.org/article/f1174d58891a4086a4fd7777cb286399
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spelling	oai:doaj.org-article:f1174d58891a4086a4fd7777cb2863992021-11-25T17:41:38ZSimple and Rapid Assembly of TALE Modules Based on the Degeneracy of the Codons and Trimer Repeats10.3390/genes121117612073-4425https://doaj.org/article/f1174d58891a4086a4fd7777cb2863992021-11-01T00:00:00Zhttps://www.mdpi.com/2073-4425/12/11/1761https://doaj.org/toc/2073-4425Transcription activator-like effectors (TALEs) have been effectively used for targeted genome editing, transcriptional regulation, epigenetic modification, and locus-specific DNA imaging. However, with the advent of the clustered regularly interspaced short palindromic repeat/Cas9 system, an easy-to-use tool with the same function as TALEs, TALEs have recently been abandoned because of their complexity, time consumption, and difficult handling in common labs. Here, we described a degenerated codon-based TALE assembly system for simple, rapid, and efficient TALE assembly. TALE trimers with nonrepetitive DNA sequences were amplified by PCR and sequentially assembled via Gibson assembly. Our method is cost-effective, requires only commonly used basic molecular biology reagents, and takes only 2 h from target sequence analysis to completion. This simple, rapid, and lab-friendly TALE assembly method will restore the value of TALEs in DNA targeting.Lingyin ChengXiaoqing ZhouYuling ZhengChengcheng TangYu LiuShuwen ZhengYang LiuJizeng ZhouChuan LiMin ChenLiangxue LaiQingjian ZouMDPI AGarticletranscription activator-like effectorsdegenerated codontrimer repeatsGibson assemblyGeneticsQH426-470ENGenes, Vol 12, Iss 1761, p 1761 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	transcription activator-like effectors degenerated codon trimer repeats Gibson assembly Genetics QH426-470
spellingShingle	transcription activator-like effectors degenerated codon trimer repeats Gibson assembly Genetics QH426-470 Lingyin Cheng Xiaoqing Zhou Yuling Zheng Chengcheng Tang Yu Liu Shuwen Zheng Yang Liu Jizeng Zhou Chuan Li Min Chen Liangxue Lai Qingjian Zou Simple and Rapid Assembly of TALE Modules Based on the Degeneracy of the Codons and Trimer Repeats
description	Transcription activator-like effectors (TALEs) have been effectively used for targeted genome editing, transcriptional regulation, epigenetic modification, and locus-specific DNA imaging. However, with the advent of the clustered regularly interspaced short palindromic repeat/Cas9 system, an easy-to-use tool with the same function as TALEs, TALEs have recently been abandoned because of their complexity, time consumption, and difficult handling in common labs. Here, we described a degenerated codon-based TALE assembly system for simple, rapid, and efficient TALE assembly. TALE trimers with nonrepetitive DNA sequences were amplified by PCR and sequentially assembled via Gibson assembly. Our method is cost-effective, requires only commonly used basic molecular biology reagents, and takes only 2 h from target sequence analysis to completion. This simple, rapid, and lab-friendly TALE assembly method will restore the value of TALEs in DNA targeting.
format	article
author	Lingyin Cheng Xiaoqing Zhou Yuling Zheng Chengcheng Tang Yu Liu Shuwen Zheng Yang Liu Jizeng Zhou Chuan Li Min Chen Liangxue Lai Qingjian Zou
author_facet	Lingyin Cheng Xiaoqing Zhou Yuling Zheng Chengcheng Tang Yu Liu Shuwen Zheng Yang Liu Jizeng Zhou Chuan Li Min Chen Liangxue Lai Qingjian Zou
author_sort	Lingyin Cheng
title	Simple and Rapid Assembly of TALE Modules Based on the Degeneracy of the Codons and Trimer Repeats
title_short	Simple and Rapid Assembly of TALE Modules Based on the Degeneracy of the Codons and Trimer Repeats
title_full	Simple and Rapid Assembly of TALE Modules Based on the Degeneracy of the Codons and Trimer Repeats
title_fullStr	Simple and Rapid Assembly of TALE Modules Based on the Degeneracy of the Codons and Trimer Repeats
title_full_unstemmed	Simple and Rapid Assembly of TALE Modules Based on the Degeneracy of the Codons and Trimer Repeats
title_sort	simple and rapid assembly of tale modules based on the degeneracy of the codons and trimer repeats
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/f1174d58891a4086a4fd7777cb286399
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Simple and Rapid Assembly of TALE Modules Based on the Degeneracy of the Codons and Trimer Repeats

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