Targeted transgene integration in plant cells using designed zinc finger nucleases View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2008-12-27

AUTHORS

Charles Q. Cai, Yannick Doyon, W. Michael Ainley, Jeffrey C. Miller, Russell C. DeKelver, Erica A. Moehle, Jeremy M. Rock, Ya-Li Lee, Robbi Garrison, Lisa Schulenberg, Ryan Blue, Andrew Worden, Lisa Baker, Farhoud Faraji, Lei Zhang, Michael C. Holmes, Edward J. Rebar, Trevor N. Collingwood, Beth Rubin-Wilson, Philip D. Gregory, Fyodor D. Urnov, Joseph F. Petolino

ABSTRACT

Targeted transgene integration in plants remains a significant technical challenge for both basic and applied research. Here it is reported that designed zinc finger nucleases (ZFNs) can drive site-directed DNA integration into transgenic and native gene loci. A dimer of designed 4-finger ZFNs enabled intra-chromosomal reconstitution of a disabled gfp reporter gene and site-specific transgene integration into chromosomal reporter loci following co-transformation of tobacco cell cultures with a donor construct comprised of sequences necessary to complement a non-functional pat herbicide resistance gene. In addition, a yeast-based assay was used to identify ZFNs capable of cleaving a native endochitinase gene. Agrobacterium delivery of a Ti plasmid harboring both the ZFNs and a donor DNA construct comprising a pat herbicide resistance gene cassette flanked by short stretches of homology to the endochitinase locus yielded up to 10% targeted, homology-directed transgene integration precisely into the ZFN cleavage site. Given that ZFNs can be designed to recognize a wide range of target sequences, these data point toward a novel approach for targeted gene addition, replacement and trait stacking in plants. More... »

PAGES

699-709

Journal

TITLE

Plant Molecular Biology

ISSUE

6

VOLUME

69

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s11103-008-9449-7

    DOI

    http://dx.doi.org/10.1007/s11103-008-9449-7

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1046854085

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/19112554


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