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Updated: Jul 3, 2026
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Genome Editing with CompoZr Custom Zinc Finger Nucleases (ZFNs)
Published on: June 14, 2012
Rapid "open-source" engineering of customized zinc-finger nucleases for highly efficient gene modification
Morgan L Maeder1, Stacey Thibodeau-Beganny, Anna Osiak
1Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA 02129, USA.
Molecular Cell
|July 29, 2008
Summary
A new method called Oligomerized Pool Engineering (OPEN) rapidly creates zinc-finger arrays for zinc-finger nucleases (ZFNs). This technology enables efficient, targeted genome modifications for research and gene therapy.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- Custom-made zinc-finger nucleases (ZFNs) offer efficient genome modification across various organisms.
- A key challenge in ZFN application is the efficient generation of specific zinc-finger arrays.
Purpose of the Study:
- To introduce Oligomerized Pool Engineering (OPEN), a rapid and open-source strategy for constructing multifinger arrays.
- To demonstrate OPEN's effectiveness compared to existing modular assembly methods.
Main Methods:
- Development and application of the OPEN strategy for engineering zinc-finger arrays.
- Construction and testing of 37 highly active ZFN pairs targeting multiple endogenous and reporter genes.
Main Results:
- OPEN proved more effective than the modular assembly method for creating zinc-finger arrays.
- The engineered ZFNs achieved high efficiencies (1%-50%) in targeted alterations at 11 distinct sites.
- Successful modifications were demonstrated in human genes (VEGF-A, HoxB13, CFTR), a plant gene (tobacco SuRA), and an EGFP reporter.
Conclusions:
- OPEN is a rapid, open-source method for engineering highly active zinc-finger arrays.
- This strategy facilitates broader application of ZFN technology in biological research and gene therapy.
