• Register
X
Forgot Password

If you have forgotten your password you can enter your email here and get a temporary password sent to your email.

X

Leaving Community

Are you sure you want to leave this community? Leaving the community will revoke any permissions you have been granted in this community.

No
Yes

FLASH assembly of TALENs for high-throughput genome editing.

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.

Pubmed ID: 22484455

Authors

  • Reyon D
  • Tsai SQ
  • Khayter C
  • Foden JA
  • Sander JD
  • Joung JK

Journal

Nature biotechnology

Publication Data

May 18, 2012

Associated Grants

  • Agency: NIGMS NIH HHS, Id: DP1 GM105378
  • Agency: NIH HHS, Id: DP1 OD006862
  • Agency: NIH HHS, Id: DP1 OD006862
  • Agency: NIH HHS, Id: DP1 OD006862-01
  • Agency: NIH HHS, Id: DP1 OD006862-02
  • Agency: NHGRI NIH HHS, Id: P50 HG005550
  • Agency: NHGRI NIH HHS, Id: P50 HG005550-01
  • Agency: NHGRI NIH HHS, Id: P50 HG005550-02
  • Agency: NHGRI NIH HHS, Id: P50HG005550
  • Agency: NCI NIH HHS, Id: T32 CA009216
  • Agency: NCI NIH HHS, Id: T32 CA009216
  • Agency: NCI NIH HHS, Id: T32 CA009216-30
  • Agency: NCI NIH HHS, Id: T32 CA009216-31

Mesh Terms

  • Automation
  • Base Sequence
  • Biotinylation
  • Computational Biology
  • DNA Restriction Enzymes
  • Endonucleases
  • Epigenesis, Genetic
  • Gene Targeting
  • Genetic Techniques
  • Genome, Human
  • Green Fluorescent Proteins
  • Humans
  • Molecular Sequence Data
  • Mutation
  • Neoplasms
  • Sequence Analysis, DNA
  • Sequence Homology, Nucleic Acid
  • Transcription, Genetic