Engineering designer transcription activator‐‐like effector nucleases (TALENs) by REAL or REAL‐fast assembly
D Reyon, C Khayter, MR Regan… - Current protocols in …, 2012 - Wiley Online Library
D Reyon, C Khayter, MR Regan, JK Joung, JD Sander
Current protocols in molecular biology, 2012•Wiley Online LibraryEngineered transcription activator–like effector nucleases (TALENs) are broadly useful tools
for performing targeted genome editing in a wide variety of organisms and cell types
including plants, zebrafish, C. elegans, rat, human somatic cells, and human pluripotent
stem cells. Here we describe detailed protocols for the serial, hierarchical assembly of
TALENs that require neither PCR nor specialized multi‐fragment ligations and that can be
implemented by any laboratory. These r estriction e nzyme a nd l igation (REAL)–based …
for performing targeted genome editing in a wide variety of organisms and cell types
including plants, zebrafish, C. elegans, rat, human somatic cells, and human pluripotent
stem cells. Here we describe detailed protocols for the serial, hierarchical assembly of
TALENs that require neither PCR nor specialized multi‐fragment ligations and that can be
implemented by any laboratory. These r estriction e nzyme a nd l igation (REAL)–based …
Abstract
Engineered transcription activator–like effector nucleases (TALENs) are broadly useful tools for performing targeted genome editing in a wide variety of organisms and cell types including plants, zebrafish, C. elegans, rat, human somatic cells, and human pluripotent stem cells. Here we describe detailed protocols for the serial, hierarchical assembly of TALENs that require neither PCR nor specialized multi‐fragment ligations and that can be implemented by any laboratory. These restriction enzyme and ligation (REAL)–based protocols can be practiced using plasmid libraries and user‐friendly, Web‐based software that both identifies target sites in sequences of interest and generates printable graphical guides that facilitate assembly of TALENs. With the described platform of reagents, protocols, and software, researchers can easily engineer multiple TALENs within 2 weeks using standard cloning techniques. Curr. Protoc. Mol. Biol. 100:12.15.1‐12.15.14. © 2012 by John Wiley & Sons, Inc.
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