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Related Experiment Video

Updated: May 23, 2026

Mouse Genome Engineering Using Designer Nucleases
12:04

Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

Precision genome engineering with programmable DNA-nicking enzymes.

Eunji Kim1, Sojung Kim, Duk Hyoung Kim

  • 1National Creative Research Initiatives Center for Genome Engineering and Department of Chemistry, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul, South Korea.

Genome Research
|April 24, 2012
PubMed
Summary

Programmable DNA-nicknicking enzymes (nickases) create single-strand breaks, enabling precise genome engineering via error-free repair. This avoids unwanted mutations common with zinc finger nucleases (ZFNs), offering broad applications in science and medicine.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Zinc finger nucleases (ZFNs) are genome engineering tools but cause random mutations via error-prone DNA repair.
  • Nonhomologous end-joining (NHEJ) repair of ZFN-induced double-strand breaks (DSBs) leads to unwanted insertions/deletions (indels).

Purpose of the Study:

  • To develop a precision genome engineering tool that avoids random mutations.
  • To present programmable DNA-nicking enzymes (nickases) as an alternative to ZFNs.

Main Methods:

  • Engineered programmable zinc finger nickases to create single-strand breaks (SSBs) instead of DSBs.
  • Leveraged error-free homologous recombination (HR) repair pathway for targeted modifications.

Main Results:

  • Zinc finger nickases induce SSBs, which are repaired by HR, not mutagenic NHEJ.
  • Site-specific genome modifications were achieved without inducing unwanted indels at on-target sites.

Conclusions:

  • Programmable nickases enable precise genome engineering by avoiding random indels.
  • Nickase technology offers broad utility in research, medicine, and biotechnology for targeted gene editing.