Phenotypic engineering by reprogramming gene transcription using novel artificial transcription factors in Escherichia coli

Ju Young Lee, Bong Hyun Sung, Byung Jo Yu, Jun Hyoung Lee, Sang Hee Lee, Mi Sun Kim, Michael D. Koob, Sun Chang Kim

Research output: Contribution to journalArticlepeer-review

49 Scopus citations

Abstract

Now that many genomes have been sequenced and the products of newly identified genes have been annotated, the next goal is to engineer the desired phenotypes in organisms of interest. For the phenotypic engineering of microorganisms, we have developed novel artificial transcription factors (ATFs) capable of reprogramming innate gene expression circuits in Escherichia coli. These ATFs are composed of zinc finger (ZF) DNA-binding proteins, with distinct specificities, fused to an E. coli cyclic AMP receptor protein (CRP). By randomly assembling 40 different types of ZFs, we have constructed more than 6.4 × 104 ATFs that consist of 3 ZF DNA-binding domains and a CRP effector domain. Using these ATFs, we induced various phenotypic changes in E. coli and selected for industrially important traits, such as resistance to heat shock, osmotic pressure and cold shock. Genes associated with the heat-shock resistance phenotype were then characterized. These results and the general applicability of this platform clearly indicate that novel ATFs are powerful tools for the phenotypic engineering of microorganisms and can facilitate microbial functional genomic studies.

Original languageEnglish (US)
Article numbere102
JournalNucleic acids research
Volume36
Issue number16
DOIs
StatePublished - 2008

Fingerprint

Dive into the research topics of 'Phenotypic engineering by reprogramming gene transcription using novel artificial transcription factors in Escherichia coli'. Together they form a unique fingerprint.

Cite this