Nucleotide kinase-based selection system for genetic switches

Kohei Ike, Daisuke Umeno

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Ever-increasing repertories of RNA-based switching devices are enabling synthetic biologists to construct compact, self-standing, and easy-to-integrate regulatory circuits. However, it is rather rare that the existing RNA-based expression controllers happen to have the exact specification needed for particular applications from the beginning. Evolutionary design of is powerful strategy for quickly tuning functions/specification of genetic switches. Presented here are the steps required for rapid and efficient enrichment of genetic switches with desired specification using recently developed nucleoside kinase-based dual selection system. Here, the library of genetic switches, created by randomizing either the part or the entire sequence coding switching components, is subjected to OFF (negative) selection and ON (positive) selection in various conditions. The entire selection process is completed only by liquid handling, facilitating the parallel and continuous operations of multiple selection projects. This automation-liable platform for genetic selection of functional switches has potential applications for development of RNA-based biosensors, expression controllers, and their integrated forms (genetic circuits).

Original languageEnglish
Title of host publicationArtificial Riboswitches
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages141-152
Number of pages12
ISBN (Print)9781627037549
DOIs
Publication statusPublished - 2014
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1111
ISSN (Print)1064-3745

Keywords

  • Directed evolution
  • Dual selection
  • Genetic selection
  • Genetic switch
  • Nucleoside analog
  • Thymidine kinase

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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