Gene Replacement in Plants

Ohl, Stephan; Offringa, Remko; Elzen, Peter J. M. van den; Hooykaas, Paul J. J.

doi:10.1007/978-94-011-1094-5_10

Homologous Recombination and Gene Silencing in Plants 1994

DOI: 10.1007/978-94-011-1094-5_10

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Gene Replacement in Plants

Stephan Ohl¹,

Remko Offringa²,

Peter J. M. van den Elzen³

et al.

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Cited by 6 publications

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“…The primary methods used in site-directed gene manipulation rely on gene replacement via homologous recombination using an appropriately designed gene targeting vector (1). In plant cells, gene targeting has been limited by the low frequency of homologous recombination (1,2).…”

mentioning

confidence: 99%

Targeted manipulation of maize genes in vivo using chimeric RNA/DNA oligonucleotides

Zhu

Peterson

Tagliani

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

189

131

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Site-specific heritable mutations in maize genes were engineered by introducing chimeric RNA͞DNA oligonucleotides. Two independent targets within the endogenous maize acetohydroxyacid synthase gene sequence were modified in a site-specific fashion, thereby conferring resistance to either imidazolinone or sulfonylurea herbicides. Similarly, an engineered green f luorescence protein transgene was site-specifically modified in vivo. Expression of the introduced inactive green f luorescence protein was restored, and plants containing the modified transgene were regenerated. Progeny analysis indicated Mendelian transmission of the converted transgene. The efficiency of gene conversion mediated by chimeric oligonucleotides in maize was estimated as 10 ؊4 , which is 1-3 orders of magnitude higher than frequencies reported for gene targeting by homologous recombination in plants. The heritable changes in maize genes engineered by this approach create opportunities for basic studies of plant gene function and agricultural trait manipulation and also provide a system for studying mismatch repair mechanisms in maize.

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