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Related Experiment Video

Updated: May 16, 2026

Genome Editing with CompoZr Custom Zinc Finger Nucleases (ZFNs)
09:11

Genome Editing with CompoZr Custom Zinc Finger Nucleases (ZFNs)

Published on: June 14, 2012

Highly active zinc-finger nucleases by extended modular assembly.

Mital S Bhakta1, Isabelle M Henry, David G Ousterout

  • 1Genome Center and Department of Biochemistry and Molecular Medicine, University of California, Davis, CA 95616, USA.

Genome Research
|December 11, 2012
PubMed
Summary

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Modular assembly (MA) of zinc-finger nucleases (ZFNs) can be inefficient. This study shows that longer, six-finger ZFN arrays significantly improve genome engineering success rates in human and mouse cells.

Area of Science:

  • Molecular Biology
  • Genome Engineering
  • Biotechnology

Background:

  • Zinc-finger nucleases (ZFNs) are crucial for genome engineering but their widespread adoption is limited by the lack of robust, noncommercial methods.
  • The modular assembly (MA) method allows rapid ZFN creation but often results in inactive nucleases, particularly with three- and four-finger arrays, with success rates below 25%.

Purpose of the Study:

  • To systematically investigate the impact of array length on ZFN activity and efficiency.
  • To identify optimal ZFN designs for enhanced genome targeting and success rates.

Main Methods:

  • Systematic study of ZFN array lengths, focusing on three- to six-finger combinations.
  • Development and application of a novel drop-out linker scheme for rapid assessment of ZFN combinations.

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Last Updated: May 16, 2026

Genome Editing with CompoZr Custom Zinc Finger Nucleases (ZFNs)
09:11

Genome Editing with CompoZr Custom Zinc Finger Nucleases (ZFNs)

Published on: June 14, 2012

Mouse Genome Engineering Using Designer Nucleases
12:04

Mouse Genome Engineering Using Designer Nucleases

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  • Analysis of 268 array variants, correlating activity with an ab initio B-score cutoff.

    • Main Results:

    • Six-finger MA ZFN arrays successfully produced mutations at 71% (15 of 21) of targeted loci in human and mouse cells.
    • Shorter arrays demonstrated improved activity in specific cases.
    • Half of MA ZFNs exceeding an ab initio B-score cutoff of 15 were active, irrespective of array composition.

    Conclusions:

    • Optimizing ZFN array length, particularly using longer six-finger arrays, significantly enhances the success rate of genome engineering.
    • The MA method, when appropriately applied with optimized array lengths, offers a highly effective approach for targeting diverse DNA sequences.
    • This research provides a pathway to more robust and successful genome engineering applications using ZFN technology.