Joung Lab TAL Effector Engineering Reagents
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The Joung Lab has developed three platforms for engineering TAL effector repeat arrays of any desired length: the REAL (Restriction Enzyme And Ligation) assembly method (Sander et al., Nat Biotechnol. 2011), the REAL-Fast assembly method (Reyon et al., manuscript submitted), and the FLASH (Fast Ligation-based Automatable Solid-phase High-throughput) assembly method (Reyon and Tsai et al., Nat Biotechnol 2012). All three of these platforms assemble TAL effector repeat arrays on a single architectural framework and the DNAs produced by all three methods are identical in sequence. TAL effector repeat arrays produced on this framework has yielded TALENs which function efficiently in C. elegans (Wood et al., Science 2011), zebrafish (Sander et al., Nat Biotechnol. 2011), rats (Tesson et al., Nat Biotechnol. 2011), human somatic (Miller et al., Nat Biotechnol. 2011; Reyon and Tsai et al., Nat Biotechnol 2012) and pluripotent stem cells (Hockemeyer et al., Nat Biotechnol. 2011 as well as TALE transcriptional activators that function efficiently in human cells (Maeder et al., Nat Methods 2013).
More detailed information about REAL, REAL-Fast and FLASH, including detailed and up-to-date protocols, can be found on the TALengineering website http://www.TALengineering.org Users may also wish to join the Genome Engineering newsgroup https://groups.google.com/group/talengineering
An upgraded version of the ZiFiT Targeter program is available to enable users to identify both dimeric TALEN and monomeric TALE transcriptional activator target sites and to simplify the construction of TALEN expression plasmids using REAL, REAL-Fast or FLASH assembly. This new version of ZiFiT Targeter enables users to search their sequences of interest for potential TALEN or TALE transcriptional activator target sites and then generates individual user-friendly graphical roadmaps for each target that guide the user through the REAL or REAL-Fast assembly process or lists of plasmid ID numbers required for FLASH assembly. The software is publicly available without registration at: http://zifit.partners.org.
Links to additional pages describing these reagents are provided below: