Rapid “Open-Source” Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification

Morgan L Maeder; Stacey Thibodeau-Beganny; Anna Osiak; David A Wright; Reshma M Anthony; Magdalena Eichtinger; Tao Jiang; Jonathan E Foley; Ronnie J Winfrey; Jeffrey A Townsend; Erica Unger-Wallace; Jeffry D Sander; Felix Müller-Lerch; Fengli Fu; Joseph Pearlberg; Carl Göbel; Justin P Dassie; Shondra M Pruett-Miller; Matthew H Porteus; Dennis C Sgroi; A. John Iafrate; Drena Dobbs; Paul B McCray; Toni Cathomen; Daniel F Voytas; J. Keith Joung

doi:10.1016/j.molcel.2008.06.016

Back

Rapid “Open-Source” Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification

Journal article

Open access

Peer reviewed

Rapid “Open-Source” Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification

Morgan L Maeder, Stacey Thibodeau-Beganny, Anna Osiak, David A Wright, Reshma M Anthony, Magdalena Eichtinger, Tao Jiang, Jonathan E Foley, Ronnie J Winfrey, Jeffrey A Townsend, …

Molecular cell, Vol.31(2), pp.294-301

2008

DOI: 10.1016/j.molcel.2008.06.016

PMCID: PMC2535758

PMID: 18657511

Files and links (1)

url

https://doi.org/10.1016/j.molcel.2008.06.016View

Published (Version of record) Open Access

Abstract

Custom-made zinc-finger nucleases (ZFNs) can induce targeted genome modifications with high efficiency in cell types including Drosophila, C. elegans, plants, and humans. A bottleneck in the application of ZFN technology has been the generation of highly specific engineered zinc-finger arrays. Here we describe OPEN (Oligomerized Pool ENgineering), a rapid, publicly available strategy for constructing multifinger arrays, which we show is more effective than the previously published modular assembly method. We used OPEN to construct 37 highly active ZFN pairs which induced targeted alterations with high efficiencies (1%–50%) at 11 different target sites located within three endogenous human genes ( VEGF-A, HoxB13, and CFTR), an endogenous plant gene (tobacco SuRA), and a chromosomally integrated EGFP reporter gene. In summary, OPEN provides an “open-source” method for rapidly engineering highly active zinc-finger arrays, thereby enabling broader practice, development, and application of ZFN technology for biological research and gene therapy.

DNA

Details

Title: Subtitle: Rapid “Open-Source” Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification
Creators: Morgan L Maeder - Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA 02129, USA
Stacey Thibodeau-Beganny - Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA 02129, USA
Anna Osiak - Institute of Virology (CBF), Charité Medical School, D-12203 Berlin, Germany
David A Wright - Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA
Reshma M Anthony - Department of Pediatrics, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Magdalena Eichtinger - Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA 02129, USA
Tao Jiang - Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA 02129, USA
Jonathan E Foley - Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA 02129, USA
Ronnie J Winfrey - Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA
Jeffrey A Townsend - Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA
Erica Unger-Wallace - Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA
Jeffry D Sander - Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA
Felix Müller-Lerch - Institute of Virology (CBF), Charité Medical School, D-12203 Berlin, Germany
Fengli Fu - Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA
Joseph Pearlberg - Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA
Carl Göbel - Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA 02129, USA
Justin P Dassie - Department of Pediatrics, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Shondra M Pruett-Miller - Department of Pediatrics, UT Southwestern Medical Center, Dallas, TX 75390, USA
Matthew H Porteus - Department of Pediatrics, UT Southwestern Medical Center, Dallas, TX 75390, USA
Dennis C Sgroi - Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA 02129, USA
A. John Iafrate - Pathology Service, Massachusetts General Hospital, Boston, MA 02114, USA
Drena Dobbs - Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA
Paul B McCray - Department of Pediatrics, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Toni Cathomen - Institute of Virology (CBF), Charité Medical School, D-12203 Berlin, Germany
Daniel F Voytas - Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA
J. Keith Joung - Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA 02129, USA
Resource Type: Journal article
Publication Details: Molecular cell, Vol.31(2), pp.294-301
Publisher: Elsevier Inc
DOI: 10.1016/j.molcel.2008.06.016
PMID: 18657511
PMCID: PMC2535758
ISSN: 1097-2765
eISSN: 1097-4164
Language: English
Date published: 2008
Academic Unit: Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Internal Medicine
Record Identifier: 9984093352602771

Metrics

12 Record Views

535 Times Cited - Web of Science

See more details