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FLASH assembly of TALENs for high-throughput genome editing
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FLASH assembly of TALENs for high-throughput genome editing
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Journal Article
FLASH assembly of TALENs for high-throughput genome editing
2012
Overview
Transcription activator–like effector nucleases (TALENs) enable genetic modification at specific sites in a genome. Reyon
et al
. present a method for high-throughput generation of TALENs, facilitating large-scale genome engineering.
Engineered transcription activator–like effector nucleases (TALENs) have shown promise as facile and broadly applicable genome editing tools. However, no publicly available high-throughput method for constructing TALENs has been published, and large-scale assessments of the success rate and targeting range of the technology remain lacking. Here we describe the fast ligation-based automatable solid-phase high-throughput (FLASH) system, a rapid and cost-effective method for large-scale assembly of TALENs. We tested 48 FLASH-assembled TALEN pairs in a human cell–based EGFP reporter system and found that all 48 possessed efficient gene-modification activities. We also used FLASH to assemble TALENs for 96 endogenous human genes implicated in cancer and/or epigenetic regulation and found that 84 pairs were able to efficiently introduce targeted alterations. Our results establish the robustness of TALEN technology and demonstrate that FLASH facilitates high-throughput genome editing at a scale not currently possible with other genome modification technologies.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ 631/337
/ Biological and medical sciences
/ Biomedical and Life Sciences
/ Biomedical Engineering/Biotechnology
/ Computational Biology - methods
/ DNA Restriction Enzymes - genetics
/ Fundamental and applied biological sciences. Psychology
/ Genomics
/ Green Fluorescent Proteins - metabolism
/ Humans
/ Molecular and cellular biology
/ Mutation
/ Proteins
/ resource
/ Sequence Analysis, DNA - instrumentation
/ Sequence Analysis, DNA - methods
