Generate TALE/TALEN as Easily and Rapidly as Generating CRISPR.

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 in vivo application. By contrast, TALE has been applied in vivo 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.

SUBMITTER: Zhang S 

PROVIDER: S-EPMC6423989 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature