Cre/lox Mediated Marker Gene Excision in Transgenic Crop Plants

Gilbertson, Larry A.; Dioh, Waly; Addae, Prince; Ekena, Joanne; Keithly, Greg E.; Neuman, Mark D.; Peschke, V. M.; Petersen, Mike; Samuelson, Chris; Subbarao, Shubha; Wei, Liping; Zhang, Wanggen; Barton, Ken

doi:10.1007/978-94-017-2679-5_43

Cited by 5 publications

(3 citation statements)

References 3 publications

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“…Additionally, its removal allows the reuse of the same selection scheme in subsequent rounds of transformation. Among the different methods, site-specific recombination has been shown effective beyond model plants (Dale and Ow 1991;Russell et al 1992), with reports of successful applications in a wide range of food crop species (Ballester et al 2006;Cao et al 2006;Cuellar et al 2006;Gilbertson et al 2003;Hoa et al 2002;Hu et al 2008;Kerbach et al 2005;Lyznik et al 1996;Radhakrishnan and Srivastava 2005;Sreekala et al 2005;Srivastava and Ow 2003;Srivastava et al 1999;Zhang et al 2003).…”

Section: Introductionmentioning

confidence: 97%

ParA resolvase catalyzes site-specific excision of DNA from the Arabidopsis genome

et al. 2008

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The small serine resolvase ParA from bacterial plasmids RK2 and RP4 catalyzes the recombination of two identical 133 bp recombination sites known as MRS. Previously, we reported that ParA is active in the fission yeast Schizosaccharomyces pombe. In this work, the parA recombinase gene was placed under the control of the Arabidopsis OXS3 promoter and introduced into Arabidopsis lines harboring a chromosomally integrated MRS-flanked target. The ParA recombinase excised the MRS-flanked DNA and the excision event was detected in subsequent generations in the absence of ParA, indicating germinal transmission of the excision event. The precise site-specific deletion by the ParA recombination system in planta demonstrates that the ParA recombinase can be used to remove transgenic DNA, such as selectable markers or other introduced transgenes that are no longer desired in the final product.

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Section: Introductionmentioning

confidence: 97%

ParA resolvase catalyzes site-specific excision of DNA from the Arabidopsis genome

et al. 2008

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“…For example, the removal of an antibiotic resistance gene eliminates the possibility of its horizontal transfer, as well permits that same selection gene to be used again for subsequent rounds of gene transfer (Dale and Ow, 1991;Russell et al, 1992). Sitespecific deletion of marker genes from major crop plants such as corn, rice, wheat, cotton, and soybean has been achieved (Gilbertson et al, 2003), and a high lysine corn line from Monsanto and Cargill, which used Cre-lox to excise away the kanamycin resistance marker, has received U.S. regulatory approval for commercial cultivation. Site-specific DNA integration also holds promise for expediting the genetic improvement of crop varieties.…”

Section: Introductionmentioning

confidence: 99%

Site‐specific recombination systems for the genetic manipulation of eukaryotic genomes

Thomson

2006

Genesis

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Site-specific recombination systems, such as the bacteriophage Cre-lox and yeast FLP-FRT systems, have become valuable tools for the rearrangement of DNA in higher eukaryotes. As a first step to expanding the repertoire of recombination tools, we screened recombination systems derived from the resolvase/invertase family for site-specific recombinase activity in the fission yeast Schizosaccharomyces pombe. Here, we report that seven recombination systems, four from the small serine resolvase subfamily (CinH, ParA, Tn1721, and Tn5053) and three from the large serine resolvase subfamily (Bxb1, TP901-1, and U153), can catalyze site-specific deletion in S. pombe. Those from the large serine resolvase subfamily were also capable of site-specific integration and inversion. In all cases, the recombination events were precise. Functional operation of these recombination systems in the fission yeast holds promise that they may be further developed as recombination tools for the site-specific rearrangement of plant and animal genomes.

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“…Removal of the selectable marker also allows reuse of the same selection regime for subsequent rounds of gene transfer. A number of recombinase-mediated marker deletion strategies have been reported in model plants Hare and Chua 2002 ;Thomson et al 2009Thomson et al , 2010Thomson et al , 2012, as well as in food crop species (Ballester et al 2007 ;Blechl et al 2012 ;Cao et al 2006 ;Chawla et al 2006 ;Chong-Pérez et al 2012 ;Cuellar et al 2006 ;Djukanovic et al 2008 ;Gilbertson et al 2003 ;Hoa et al 2002 ;Hu et al 2008 ;Kapusi et al 2012 ;Kempe et al 2010 ;Kerbach et al 2005 ;Lyznik et al 1996 ;Radhakrishnan and Srivastava 2005 ;Rubtsova et al 2008 ;Sreekala et al 2005 ;Srivastava et al 1999 ;Srivastava and Ow 2003 ;Wurdig et al 2013 ;Zhang et al 2003 ;Zou et al 2013 ). Recent reviews on the topic of recombinase-mediated excision strategies in plants are Gidoni et al ( 2008 ), Moon et al ( 2011 ), andWang et al ( 2011 ).…”

Section: Application: Excisionmentioning

confidence: 99%