FLP-mediated recombination of FRT sites in the maize genome

Lyznik, L. Alexander; Rao, K. V.; Hodges, Thomas K.

doi:10.1093/nar/24.19.3784

Cited by 104 publications

(55 citation statements)

References 42 publications

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“…Several methods have been proposed to eliminate the selection marker from the primary transformant. These include use of a site-specific recombination system, such as Cre-lox or Flp-Frt (2,19,57,209,235,347,348) to remove the marker, transposon-based movement of the selection marker from the initial site of insertion from the plant genome entirely or to another unlinked site from which it can be segregated in subsequent generations (93), or the use of multiple T-DNAs which can insert into unlinked sites for future segregation (reviewed in references 142 and 372). Each of these systems has advantages and disadvantages.…”

Section: Transfer Of Multiple T-dnas Into the Same Plant Cell And Gementioning

confidence: 99%

Agrobacterium -Mediated Plant Transformation: the Biology behind the “Gene-Jockeying” Tool

Gelvin

2003

Microbiol Mol Biol Rev

1,139

687

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SUMMARY Agrobacterium tumefaciens and related Agrobacterium species have been known as plant pathogens since the beginning of the 20th century. However, only in the past two decades has the ability of Agrobacterium to transfer DNA to plant cells been harnessed for the purposes of plant genetic engineering. Since the initial reports in the early 1980s using Agrobacterium to generate transgenic plants, scientists have attempted to improve this “natural genetic engineer” for biotechnology purposes. Some of these modifications have resulted in extending the host range of the bacterium to economically important crop species. However, in most instances, major improvements involved alterations in plant tissue culture transformation and regeneration conditions rather than manipulation of bacterial or host genes. Agrobacterium-mediated plant transformation is a highly complex and evolved process involving genetic determinants of both the bacterium and the host plant cell. In this article, I review some of the basic biology concerned with Agrobacterium-mediated genetic transformation. Knowledge of fundamental biological principles embracing both the host and the pathogen have been and will continue to be key to extending the utility of Agrobacterium for genetic engineering purposes.

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Section: Transfer Of Multiple T-dnas Into the Same Plant Cell And Gementioning

confidence: 99%

Agrobacterium -Mediated Plant Transformation: the Biology behind the “Gene-Jockeying” Tool

Gelvin

2003

Microbiol Mol Biol Rev

1,139

687

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“…For example, marker genes can be efficiently removed by directly flanking the marker gene expression cassette with recombination sites, such as lox and FRT (Dale and Ow 1991;Lyznik et al 1996) as one method among other (Komari et al 1996) to make marker free transgenic crops. It has also been used for gene targeting to insert foreign genes precisely into a pre-existing site for more predictable expression of introduced genes than occurs with random integrations (Day et al 2000).…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium-mediated transformation of maize (Zea mays) with Cre-lox site specific recombination cassettes in BIBAC vectors

Vega

Han

et al. 2008

Plant Mol Biol

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The Cre/loxP site-specific recombination system has been applied in various plant species including maize (Zea mays) for marker gene removal, gene targeting, and functional genomics. A BIBAC vector system was adapted for maize transformation with a large fragment of genetic material including a herbicide resistance marker gene, a 30 kb yeast genomic fragment as a marker for fluorescence in situ hybridization (FISH), and a 35S-loxcre recombination cassette. Seventy-five transgenic lines were generated from Agrobacterium-mediated transformation of a maize Hi II line with multiple B chromosomes. Eighty-four inserts have been localized among all 10 A chromosome pairs by FISH using the yeast DNA probe together with a karyotyping cocktail. No inserts were found on the B chromosomes; thus a bias against the B chromosomes by the Agrobacterium-mediated transformation was revealed. The expression of a cre gene was confirmed in 68 of the 75 transgenic lines by a reporter construct for cre/lox mediated recombination. The placement of the cre/lox site-specific recombination system in many locations in the maize genome will be valuable materials for gene targeting and chromosome engineering.

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“…The latter is often referred to as auto-excision. In the first subcategory, the recombinase can be delivered to a transgenic plant by retransformation (Odell et al, 1990;Dale and Ow, 1991;Lyznik et al, 1996), by sexual crosses (Bayley et al, 1992;Russell et al, 1992;Kilby et al, 1995;Kerbach et al, 2005), or by transiently expressing the recombinase (Gleave et al, 1999;Kopertekh et al, 2004aKopertekh et al, , 2004bKopertekh and Schliemann, 2005;Jia et al, 2006). Recently, the company Renessen received U.S. regulatory approval (http://www.aphis.usda.gov/ brs/aphisdocs2 /04_22901p_com.pdf) for the transgenic line LY038 from which the selectable marker, originally present between tandemly oriented lox sites, was removed through introduction of the cre gene by a sexual cross (Ow, 2007).…”

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confidence: 99%

Marker-Free Transgenic Plants through Genetically Programmed Auto-Excision

et al. 2007

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We present here a vector system to obtain homozygous marker-free transgenic plants without the need of extra handling and within the same time frame as compared to transformation methods in which the marker is not removed. By introducing a germline-specific auto-excision vector containing a cre recombinase gene under the control of a germline-specific promoter, transgenic plants become genetically programmed to lose the marker when its presence is no longer required (i.e. after the initial selection of primary transformants). Using promoters with different germline functionality, two modules of this genetic program were developed. In the first module, the promoter, placed upstream of the cre gene, confers CRE functionality in both the male and the female germline or in the common germline (e.g. floral meristem cells). In the second module, a promoter conferring single germline-specific CRE functionality was introduced upstream of the cre gene. Promoter sequences used in this work are derived from the APETALA1 and SOLO DANCERS genes from Arabidopsis (Arabidopsis thaliana) Columbia-0 conferring common germline and single germline functionality, respectively. Introduction of the genetic program did not reduce transformation efficiency. Marker-free homozygous progeny plants were efficiently obtained, regardless of which promoter was used. In addition, simplification of complex transgene loci was observed.

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FLP-mediated recombination of FRT sites in the maize genome

Cited by 104 publications

References 42 publications

Agrobacterium -Mediated Plant Transformation: the Biology behind the “Gene-Jockeying” Tool

Agrobacterium -Mediated Plant Transformation: the Biology behind the “Gene-Jockeying” Tool

Agrobacterium-mediated transformation of maize (Zea mays) with Cre-lox site specific recombination cassettes in BIBAC vectors

Marker-Free Transgenic Plants through Genetically Programmed Auto-Excision

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