Site-specific recombination for genetic engineering in plants

Lyznik, L. Alexander; Gordon‐Kamm, William; Tao, Yumin

doi:10.1007/s00299-003-0616-7

Cited by 58 publications

(39 citation statements)

References 67 publications

(69 reference statements)

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“…First, they have been successfully employed for the removal of selectable markers from transgenic plants (Hoa et al, 2002;Zhang et al, 2003;Srivastava and Ow, 2004;Sreekala et al, 2005;Wang et al, 2005;Jia et al, 2006;Moore and Srivastava, 2006), and, second, they can be used to precisely integrate single-copy transgenes into the plant genome (Albert et al, 1995;Day et al, 2000;Srivastava and Ow, 2002;Lyznik et al, 2003;Chawla et al, 2006). This latter approach requires two rounds of transformation: in the first, a target site, such as the loxP site, is randomly introduced into the plant genome, and, in the second, a loxP-containing DNA construct is integrated into this genomic target site.…”

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confidence: 99%

Generation of Single-Copy T-DNA Transformants in Arabidopsis by the CRE/loxP Recombination-Mediated Resolution System

et al. 2007

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We investigated whether complex T-DNA loci, often resulting in low transgene expression, can be resolved efficiently into single copies by CRE/loxP-mediated recombination. An SB-loxP T-DNA, containing two invertedly oriented loxP sequences located inside and immediately adjacent to the T-DNA border ends, was constructed. Regardless of the orientation and number of SB-loxP-derived T-DNAs integrated at one locus, recombination between the outermost loxP sequences in direct orientation should resolve multiple copies into a single T-DNA copy. Seven transformants with a complex SB-loxP locus were crossed with a CRE-expressing plant. In three hybrids, the complex T-DNA locus was reduced efficiently to a single-copy locus. Upon segregation of the CRE recombinase gene, only the simplified T-DNA locus was found in the progeny, demonstrating DNA had been excised efficiently in the progenitor cells of the gametes. In the two transformants with an inverted T-DNA repeat, the T-DNA resolution was accompanied by at least a 10-fold enhanced transgene expression. Therefore, the resolution of complex loci to a single-copy T-DNA insert by the CRE/loxP recombination system can become a valuable method for the production of elite transgenic Arabidopsis thaliana plants that are less prone to gene silencing.

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confidence: 99%

Generation of Single-Copy T-DNA Transformants in Arabidopsis by the CRE/loxP Recombination-Mediated Resolution System

et al. 2007

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“…Insertion of the cloned DNA into the chromosome can avoid these problems; however, current methods of cloned DNA insertion for use with Methanosarcina are less efficient by a factor of about 100 than transformation with autonomous plasmids because of their dependence on homologous recombination. In other organisms, methods utilizing site-specific recombination, instead of homologous recombination, have allowed much higher integration efficiencies (e.g., Lyznik et al 2003, Schweizer 2003, and references therein). One particularly useful site-specific recombinase system utilizes the Streptomyces bacteriophage φC31 integrase (Thorpe and Smith 1998).…”

Section: Introductionmentioning

confidence: 99%

New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species

Guss

Rother

Zhang

et al. 2008

Archaea

123

179

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Summary A highly efficient method for chromosomal integration of cloned DNA into Methanosarcina spp. was developed utilizing the site-specific recombination system from the Streptomyces phage φC31. Host strains expressing the φC31 integrase gene and carrying an appropriate recombination site can be transformed with non-replicating plasmids carrying the complementary recombination site at efficiencies similar to those obtained with self-replicating vectors. We have also constructed a series of hybrid promoters that combine the highly expressed M. barkeri PmcrB promoter with binding sites for the tetracycline-responsive, bacterial TetR protein. These promoters are tightly regulated by the presence or absence of tetracycline in strains that express the tetR gene. The hybrid promoters can be used in genetic experiments to test gene essentiality by placing a gene of interest under their control. Thus, growth of strains with tetR-regulated essential genes becomes tetracycline-dependent. A series of plasmid vectors that utilize the site-specific recombination system for construction of reporter gene fusions and for tetracycline regulated expression of cloned genes are reported. These vectors were used to test the efficiency of translation at a variety of start codons. Fusions using an ATG start site were the most active, whereas those using GTG and TTG were approximately one half or one fourth as active, respectively. The CTG fusion was 95% less active than the ATG fusion.

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“…Many other effective recombination systems are well established in the GT of mouse and random transformation in higher plants in addition to Cre-loxP, such as R-RS, FLP-FRT, and fC31-att (Odell and Russell 1994b;Ebinuma et al 2001;Hare and Chua 2001;Ow 2002;Lyznik et al 2003). Simultaneously, it has been realized that our strong positive-negative selection-mediated GT in rice is quite effective and widely applicable to any gene region without any other random integration (Terada et al 2002;Iida and Terada 2004; on the basis of acumulative information of the whole genome sequence (IRGSP 2005).…”

Section: Discussionmentioning

confidence: 99%

“…Such undesirable recombination events were reported in the gene-specific selection procedures described above (Hanin et al 2001;Endo et al 2007). Therefore, the combined use of site-specific recombination with our strong positive-negative selection is expected to create various gene modifications at the targeted gene locus as in mouse conditional GT (Brian 1998;Gu et al 1994;Lyznik et al 2003).…”

mentioning

confidence: 99%

Cre-loxP mediated marker elimination and gene reactivation at the waxy locus created in rice genome based on strong positive-negative selection

Terada

Nagahara

Furukawa

et al. 2010

Plant Biotechnology

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Site-specific recombination for genetic engineering in plants

Cited by 58 publications

References 67 publications

Generation of Single-Copy T-DNA Transformants in Arabidopsis by the CRE/loxP Recombination-Mediated Resolution System

Generation of Single-Copy T-DNA Transformants in Arabidopsis by the CRE/loxP Recombination-Mediated Resolution System

New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species

Cre-loxP mediated marker elimination and gene reactivation at the waxy locus created in rice genome based on strong positive-negative selection

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