2016
DOI: 10.1089/hum.2015.172
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FusX: A Rapid One-Step Transcription Activator-Like Effector Assembly System for Genome Science

Abstract: Transcription activator-like effectors (TALEs) are extremely effective, single-molecule DNA-targeting molecular cursors used for locus-specific genome science applications, including high-precision molecular medicine and other genome engineering applications. TALEs are used in genome engineering for locus-specific DNA editing and imaging, as artificial transcriptional activators and repressors, and for targeted epigenetic modification. TALEs as nucleases (TALENs) are effective editing tools and offer high bind… Show more

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Cited by 48 publications
(51 citation statements)
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References 52 publications
(58 reference statements)
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“…The Ekker lab takes scalable assembly of TALENs a step further, reducing the hands-on time for construction of new reagents to a single-day, single-tube reaction via the FusX system. 2 Furthermore, the FusX method of TALEN construction can be automated using liquid handling robots and simple inventory management programming that won't break the laboratory budget.…”
mentioning
confidence: 99%
“…The Ekker lab takes scalable assembly of TALENs a step further, reducing the hands-on time for construction of new reagents to a single-day, single-tube reaction via the FusX system. 2 Furthermore, the FusX method of TALEN construction can be automated using liquid handling robots and simple inventory management programming that won't break the laboratory budget.…”
mentioning
confidence: 99%
“…TALEN constructs were generated using the FusX kit (Addgene # 1000000063) as previously described(30). In short, RCIscript-GoldyTALEN was linearized with BsmBI (NEB R0580.…”
Section: Methodsmentioning
confidence: 99%
“…One option for mtDNA targeting includes CRISPR/Cas9; however, targeting to mitochondria would require effective gRNA importation, a hurdle yet unresolved 1,17 . TALE proteins are traditionally more difficult to assemble than CRISPR/Cas9, but single-reaction synthesis technology makes TALEs readily accessible for genome engineering applications 18 . Importantly, TALE use in mtDNA engineering only require a single mitochondrial targeting sequence (S Figure 1) that was developed from an in vivo protein trapping method 19 .…”
Section: Textmentioning
confidence: 99%
“…The final mitoTALE backbone plasmids used in this paper will be made available at Addgene. To target a specific sequences in the genome, RVDs were cloned into the GoldyTALE backbone using the FusX assembly system 18 . The RVDs used were HD=C, NN=G, NI=A, NG=T.…”
Section: Mitotale-nickase and -Nuclease Assemblymentioning
confidence: 99%