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Tailor-Made Mutations in Arabidopsis Using Zinc Finger Nucleases

  • Yiping Qi
  • Colby G. Starker
  • Feng Zhang
  • Nicholas J. Baltes
  • Daniel F. Voytas
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1062)

Abstract

Zinc finger nucleases (ZFNs) are proteins engineered to make site-specific double-strand breaks (DSBs) in a DNA sequence of interest. Imprecise repair of the ZFN-induced DSBs by the nonhomologous end-joining (NHEJ) pathway results in a spectrum of mutations, such as nucleotide substitutions, insertions, and deletions. Here we describe a method for targeted mutagenesis in Arabidopsis with ZFNs, which are engineered by context-dependent assembly (CoDA). This ZFN-induced mutagenesis method is an alternative to other currently available gene knockout or knockdown technologies and is useful for reverse genetic studies.

Key words

Arabidopsis ZFN NHEJ CoDA Mutagenesis 

Notes

Acknowledgments

This work is supported by grants from the National Science Foundation to D.F.V. (DBI 0923827 and MCB 0209818).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Yiping Qi
    • 1
  • Colby G. Starker
    • 1
  • Feng Zhang
    • 2
  • Nicholas J. Baltes
    • 1
  • Daniel F. Voytas
    • 3
  1. 1.Department of Genetics, Cell Biology & Development and Center for Genome EngineeringUniversity of MinnesotaMinneapolisUSA
  2. 2.Cellectic Plant SciencesSt. PaulUSA
  3. 3.Department of Genetics, Cell Biology and Development, Center for Genome EngineeringUniversity of MinnesotaMinneapolisUSA

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