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 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

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 P. Dassie, Shondra M. Pruett-Miller, Matthew H. Porteus, Dennis C. SgroiA. John Iafrate, Drena Dobbs, Paul B. McCray, Toni Cathomen, Daniel F. Voytas, J. Keith Joung

Research output: Contribution to journal › Article › peer-review

603 Scopus citations

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.

Original language	English (US)
Pages (from-to)	294-301
Number of pages	8
Journal	Molecular Cell
Volume	31
Issue number	2
DOIs	https://doi.org/10.1016/j.molcel.2008.06.016
State	Published - Jul 25 2008

Bibliographical note

Funding Information:
We thank Chong Jin Park for help with statistical calculations and Andrew Hirsh for helpful discussions and comments. J.K.J. is supported by the NIH (R01GM069906, R24GM078369, and R21RR024189), the Cystic Fibrosis Foundation (CFF) (MCCRAY07G0), and the MGH Pathology Service. D.F.V. is supported by the NSF (DBI 0501678). T.C. is supported by the European Commission's 6th Framework Programme (037783 ZNIP). P.B.M. is supported by the CFF (MCCRAY07G0) and the Roy J. Carver Charitable Trust. A.J.I. is supported by the MGH Pathology Service. D.C.S. is supported by the NIH (R01CA112021 and NCI SPORE in Breast Cancer at MGH). M.H.P. is supported by the NIH (R01 HL079295). J.K.J. dedicates this paper to the memory of Robert L. Burghoff and his belief in the power of methods.

Keywords

DNA

Access

10.1016/j.molcel.2008.06.016

OpenUrl availability

Full text

Cite this

Maeder, M. L., Thibodeau-Beganny, S., Osiak, A., Wright, D. A., Anthony, R. M., Eichtinger, M., Jiang, T., Foley, J. E., Winfrey, R. J., Townsend, J. A., Unger-Wallace, E., Sander, J. D., Müller-Lerch, F., Fu, F., Pearlberg, J., Göbel, C., Dassie, JP. P., Pruett-Miller, S. M., Porteus, M. H., ... Joung, J. K. (2008). Rapid "Open-Source" Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification. Molecular Cell, 31(2), 294-301. https://doi.org/10.1016/j.molcel.2008.06.016

Maeder, ML, Thibodeau-Beganny, S, Osiak, A, Wright, DA, Anthony, RM, Eichtinger, M, Jiang, T, Foley, JE, Winfrey, RJ, Townsend, JA, Unger-Wallace, E, Sander, JD, Müller-Lerch, F, Fu, F, Pearlberg, J, Göbel, C, Dassie, JPP, Pruett-Miller, SM, Porteus, MH, Sgroi, DC, Iafrate, AJ, Dobbs, D, McCray, PB, Cathomen, T, Voytas, DF & Joung, JK 2008, 'Rapid "Open-Source" Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification', Molecular Cell, vol. 31, no. 2, pp. 294-301. https://doi.org/10.1016/j.molcel.2008.06.016

@article{c2cd8bc7e7c549b487a9afeb66a449c8,

title = "Rapid {"}Open-Source{"} Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification",

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.",

keywords = "DNA",

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

note = "Funding Information: We thank Chong Jin Park for help with statistical calculations and Andrew Hirsh for helpful discussions and comments. J.K.J. is supported by the NIH (R01GM069906, R24GM078369, and R21RR024189), the Cystic Fibrosis Foundation (CFF) (MCCRAY07G0), and the MGH Pathology Service. D.F.V. is supported by the NSF (DBI 0501678). T.C. is supported by the European Commission's 6th Framework Programme (037783 ZNIP). P.B.M. is supported by the CFF (MCCRAY07G0) and the Roy J. Carver Charitable Trust. A.J.I. is supported by the MGH Pathology Service. D.C.S. is supported by the NIH (R01CA112021 and NCI SPORE in Breast Cancer at MGH). M.H.P. is supported by the NIH (R01 HL079295). J.K.J. dedicates this paper to the memory of Robert L. Burghoff and his belief in the power of methods. ",

year = "2008",

month = jul,

day = "25",

doi = "10.1016/j.molcel.2008.06.016",

language = "English (US)",

volume = "31",

pages = "294--301",

journal = "Molecular Cell",

issn = "1097-2765",

publisher = "Cell Press",

number = "2",

}

TY - JOUR

T1 - Rapid "Open-Source" Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification

AU - Maeder, Morgan L.

AU - Thibodeau-Beganny, Stacey

AU - Osiak, Anna

AU - Wright, David A.

AU - Anthony, Reshma M.

AU - Eichtinger, Magdalena

AU - Jiang, Tao

AU - Foley, Jonathan E.

AU - Winfrey, Ronnie J.

AU - Townsend, Jeffrey A.

AU - Unger-Wallace, Erica

AU - Sander, Jeffry D.

AU - Müller-Lerch, Felix

AU - Fu, Fengli

AU - Pearlberg, Joseph

AU - Göbel, Carl

AU - Dassie, Justin P P.

AU - Pruett-Miller, Shondra M.

AU - Porteus, Matthew H.

AU - Sgroi, Dennis C.

AU - Iafrate, A. John

AU - Dobbs, Drena

AU - McCray, Paul B.

AU - Cathomen, Toni

AU - Voytas, Daniel F.

AU - Joung, J. Keith

N1 - Funding Information: We thank Chong Jin Park for help with statistical calculations and Andrew Hirsh for helpful discussions and comments. J.K.J. is supported by the NIH (R01GM069906, R24GM078369, and R21RR024189), the Cystic Fibrosis Foundation (CFF) (MCCRAY07G0), and the MGH Pathology Service. D.F.V. is supported by the NSF (DBI 0501678). T.C. is supported by the European Commission's 6th Framework Programme (037783 ZNIP). P.B.M. is supported by the CFF (MCCRAY07G0) and the Roy J. Carver Charitable Trust. A.J.I. is supported by the MGH Pathology Service. D.C.S. is supported by the NIH (R01CA112021 and NCI SPORE in Breast Cancer at MGH). M.H.P. is supported by the NIH (R01 HL079295). J.K.J. dedicates this paper to the memory of Robert L. Burghoff and his belief in the power of methods.

PY - 2008/7/25

Y1 - 2008/7/25

N2 - 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.

AB - 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.

KW - DNA

UR - http://www.scopus.com/inward/record.url?scp=47349097567&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=47349097567&partnerID=8YFLogxK

U2 - 10.1016/j.molcel.2008.06.016

DO - 10.1016/j.molcel.2008.06.016

M3 - Article

C2 - 18657511

AN - SCOPUS:47349097567

SN - 1097-2765

VL - 31

SP - 294

EP - 301

JO - Molecular Cell

JF - Molecular Cell

IS - 2

ER -

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

Abstract

Bibliographical note

Keywords

Access

OpenUrl availability

Other files and links

Fingerprint

Cite this