Highly efficient genome editing via 2A-coupled co-expression of two TALEN monomersReportar como inadecuado




Highly efficient genome editing via 2A-coupled co-expression of two TALEN monomers - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

BMC Research Notes

, 7:628

First Online: 10 September 2014Received: 20 November 2013Accepted: 04 September 2014DOI: 10.1186-1756-0500-7-628

Cite this article as: Mariano, A., Xu, L. & Han, R. BMC Res Notes 2014 7: 628. doi:10.1186-1756-0500-7-628

Abstract

BackgroundTranscription activator-like effector nucleases TALENs are a useful tool for targeted gene editing. TALEN monomers are traditionally expressed from two different plasmids. Each encodes a different TALEN arm that binds to a user-defined sequence and mediates gene editing. Expression of TALEN monomers in two separate plasmids requires co-delivery of each plasmid to the cell. Efficacy of gene editing may be increased if each monomer was transcribed from the same reading frame.

FindingsWe developed a TALEN scaffold which expresses both TALEN monomers from a single open reading frame in equal molar amount by linking both monomers with a 2A self-cleaving peptide sequence. This TALEN scaffold, named pTAL10, demonstrates higher levels of genome editing than co-transfected TALENs at similar levels of transfection efficiencies when analyzed for TALEN-induced small insertions and deletions.

ConclusionsThis protocol for gene editing using 2A-linked TALENs requires transfection of only one plasmid as compared to transfection of two separate plasmids encoding each TALEN monomers.

KeywordsGene editing TALEN 2A self-cleaving sequence AbbreviationsPCRPolymerase chain reaction

TALENTranscription activator-like effector nuclease

GFPGreen fluorescent protein

RVDRepeat-variable di-residue

MSTNMyostatin

HAHemagglutinin

T7E1T7 endonuclease I.

Electronic supplementary materialThe online version of this article doi:10.1186-1756-0500-7-628 contains supplementary material, which is available to authorized users.

Download fulltext PDF



Autor: Andrew Mariano - Li Xu - Renzhi Han

Fuente: https://link.springer.com/







Documentos relacionados