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Updated: May 28, 2026

Mouse Genome Engineering Using Designer Nucleases
12:04

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Published on: April 2, 2014

Engineering designer nucleases with customized cleavage specificities.

Jeffry D Sander1, Morgan L Maeder, J Keith Joung

  • 1Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.

Current Protocols in Molecular Biology
|October 12, 2011
PubMed
Summary
This summary is machine-generated.

This study details protocols for engineering zinc finger nucleases (ZFNs) to precisely modify genomic sequences. These engineered nucleases offer efficient gene editing across diverse organisms and cell types.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Engineered designer nucleases enable precise genomic sequence modification.
  • Zinc finger nucleases (ZFNs) are widely utilized for gene editing.
  • ZFNs comprise an engineered zinc finger array fused to a cleavage domain.

Purpose of the Study:

  • To describe protocols for engineering ZFNs.
  • To enable targeted modification of specific gene sequences.
  • To facilitate ZFN application in various model organisms and cell types.

Main Methods:

  • Engineering of zinc finger arrays.
  • Fusion to a non-specific cleavage domain.
  • Utilizing the context-dependent assembly (CoDA) method for ZFN construction.

Main Results:

  • Development of protocols for ZFN engineering.
  • Demonstration of ZFNs targeting specific endogenous genes.
  • Successful application in diverse model organisms and cell types.

Conclusions:

  • Protocols for engineering ZFNs using the CoDA method are established.
  • ZFNs provide an efficient tool for genomic modification.
  • This work supports the broad application of ZFNs in biological research.