Fully Automated One-Step Synthesis of Single-Transcript TALEN Pairs Using a Biological Foundry

Ran Chao, Jing Liang, Ipek Tasan, Tong Si, Linyang Ju, Huimin Zhao

Research output: Contribution to journalArticle

Abstract

Transcription activator-like effector nuclease (TALEN) is a programmable genome editing tool with wide applications. Since TALENs perform cleavage of DNA as heterodimers, a pair of TALENs must be synthesized for each target genome locus. Conventionally, TALEN pairs are either expressed on separate vectors or synthesized separately and then subcloned to the same vector. Neither approach allows high-throughput construction of TALEN libraries for large-scale applications. Here we present a single-step assembly scheme to synthesize and express a pair of TALENs in a single-transcript format with the help of a P2A self-cleavage sequence. Furthermore, we developed a fully automated platform to custom manufacture TALENs in a versatile biological foundry. 400 pairs of TALENs can be synthesized with over 96.2% success rate at a material cost of $2.1/pair. This platform opens the door to TALEN-based genome-wide studies.

Original languageEnglish (US)
Pages (from-to)678-685
Number of pages8
JournalACS synthetic biology
Volume6
Issue number4
DOIs
StatePublished - Apr 21 2017

Keywords

  • DNA assembly
  • TALEN
  • automation
  • biofoundry
  • genetic screening
  • genome engineering

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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