TALENs, fusion proteins of DNA binding domains of TAL (transcription activator-like) effectors and the DNA cleavage domains of endonuclease FokI, have emerged as genetic tools for targeted gene modification, holding great potential for basic and applied research, even for gene therapy. Here we present a simple and efficient approach to custom-engineering TALEN genes with four basic TAL repeats and their DNA recognition cipher. The “modular assembly” method also involves the “Golden Gate” cloning strategy, using 53 ready-to-use plasmids in just two rounds of restriction and ligation to assemble TALENs with up to 24 repeat units that recognize up to 24 bp of target DNA.
