Generate TALE/TALEN As Easily and Rapidly As Generating CRISPR

Overview

Journal Mol Ther Methods Clin Dev

Publisher Cell Press

Date 2019 Mar 30

PMID 30923728

Citations 7

Authors

Shuyan Zhang

Huiting Chen

Jinke Wang

Affiliations

Soon will be listed here.

Abstract

TALE has always had potential as a gene-editing and regulatory tool. However, with the advent of CRISPR/Cas9, an easier to use tool with the same function, TALE has recently been abandoned because of the time-consuming and low-efficiency process required for its construction. The off-target activity of CRISPR/Cas9 has been a challenge to its application. By contrast, TALE has been applied for gene editing and therapy because of its high targeting capability. To overcome the key limitation of the TALE technique, we developed a high-efficiency method for constructing custom TALEs. We created 62 new monomers and developed a new pipeline that enabled assembly of custom TALEs in just 1 day. We verified the new method by assembling nine TALEs targeting the promoters of two transcription factor genes: HNF4α and E47. The expression of the two endogenous genes in two cancer cells, HepG2 and PANC1, was activated by the constructed TALEs, which promoted differentiation of the two cancer cells. Using the new method, custom TALEs can be generated as easily and rapidly as CRISPR, thus promoting the wide application of TALE-based techniques.

Citing Articles

In search of an ideal template for therapeutic genome editing: A review of current developments for structure optimization.

Shakirova A, Karpov T, Komarova Y, Lepik K Front Genome Ed. 2023; 5:1068637.

PMID: 36911237 PMC: 9992834. DOI: 10.3389/fgeed.2023.1068637.

Gene regulatory and gene editing tools and their applications for retinal diseases and neuroprotection: From proof-of-concept to clinical trial.

Altay H, Ozdemir F, Afghah F, Kilinc Z, Ahmadian M, Tschopp M Front Neurosci. 2022; 16:924917.

PMID: 36340792 PMC: 9630553. DOI: 10.3389/fnins.2022.924917.

TALE.Sense: A Versatile DNA Sensor Platform for Live Mammalian Cells.

Taghbalout A, Jillette N, Cheng A ACS Synth Biol. 2021; 11(1):116-124.

PMID: 34931802 PMC: 9202581. DOI: 10.1021/acssynbio.1c00212.

Comparison of the Feasibility, Efficiency, and Safety of Genome Editing Technologies.

Gonzalez Castro N, Bjelic J, Malhotra G, Huang C, Alsaffar S Int J Mol Sci. 2021; 22(19).

PMID: 34638696 PMC: 8509008. DOI: 10.3390/ijms221910355.

One-Day TALEN Assembly Protocol and a Dual-Tagging System for Genome Editing.

Zhang S, Wang J, Wang J ACS Omega. 2020; 5(31):19702-19714.

PMID: 32803065 PMC: 7424704. DOI: 10.1021/acsomega.0c02396.

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