title: Highly efficient genome editing via 2A-coupled co-expression of two TALEN monomers.
abstract: BACKGROUND
  Transcription 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.
  
  
  RESULTS
  We 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.
  
  
  CONCLUSIONS
  This 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.
authors.forename: Andrew
authors.initials: A
authors.lastname: Mariano
authors.forename: Li
authors.initials: L
authors.lastname: Xu
authors.affiliation: Department of Surgery, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA. renzhi.han@osumc.edu.
authors.forename: Renzhi
authors.initials: R
authors.lastname: Han
mesh_terms: D004720:Endonucleases / Q000378:metabolism*; D015894:Genome, Human; D006801:Humans; D017393:RNA Editing; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D014157:Transcription Factors; D004720:Endonucleases