Transposase is the only nematode protein required for in vitro transposition of Tc1.

Vos, J. Chris; Baere, Ivo De; Plasterk, Ronald H.A.

doi:10.1101/gad.10.6.755

Cited by 176 publications

(144 citation statements)

References 37 publications

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“…Manifestation of such a wide distribution leads to the inference that elements of this family require no or few factors produced from host cells for their transposition reaction. This has already been demonstrated with mariner, Tc1 and some other elements: transposition occurs in vitro as long as a purified transposase is provided (Vos et al, 1996;Lampe et al, 1996). It was also inferred and then proven that elements of this family move when they are artificially introduced into organisms other than their original host species.…”

Section: Introductionmentioning

confidence: 81%

The Tol1 element of the medaka fish, a member of the hAT transposable element family, jumps in Caenorhabditis elegans

Takagi

Koga

2008

Heredity

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Tol1 is a DNA-based transposable element residing in the genome of the medaka fish Oryzias latipes, and has been proven to be transposed in various vertebrate species, including mammals. This element belongs to the hAT (hobo/ Activator/Tam3) transposable element family, whose members are distributed in a wide range of organisms. It is thus possible that Tol1 is mobile in organisms other than vertebrates. We here show that transposition of this element occurs in the nematode Caenorhabditis elegans. A donor plasmid containing a Tol1 element and a helper plasmid carrying the transposase gene were delivered into gonad cells and, after several generations of culturing, were recovered from worms. PCR analysis of the donor plasmid, using primers that encompassed the Tol1 element, revealed excision of the Tol1 portion from the plasmid. Analysis of genomic DNA of the worms by the inverse PCR method provided evidence that Tol1 had been integrated into the C. elegans chromosomes. Vertebrates and C. elegans are phylogenetically distantly related organisms in that the former are deuterostomes and the latter a protostome animal. Our results indicate (1) the transposition reaction of the Tol1 element requires, besides the transposase, no factors from host cells, or (2) the host factors, even if required, are those that are common to protostomes and deuterostomes. The results also have significance for the development of a gene transfer vector and other biotechnology tools for C. elegans.

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Section: Introductionmentioning

confidence: 81%

The Tol1 element of the medaka fish, a member of the hAT transposable element family, jumps in Caenorhabditis elegans

Takagi

Koga

2008

Heredity

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“…Unlike P-element, members of the Tc1͞mariner superfamily of transposable elements do not require host-specific factors for their activity in vitro (1,2). Indeed, several of these cut-and-paste transposons were shown to be active in a wide variety of species other than their original hosts, including bacteria [Himar1 (3)], malaria mosquito [Minos (4)], zebrafish [Tc3 (5) and Mos1 (6)], chicken [Mos1 (7)], and human cells [Tc1 (8), Sleeping Beauty (SB) (9), Himar1 (10), and Minos (11)].…”

Section: T Ransposon Tagging Is a Valuable Tool For Functional Genomimentioning

confidence: 99%

Regulated transposition of a fish transposon in the mouse germ line

Fischer

Wienholds

Plasterk

2001

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

212

187

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Tc1͞mariner elements are able to transpose in species other than the host from which they were isolated. As potential vectors for insertional mutagenesis and transgenesis of the mouse, these cut-and-paste transposons were tested for their ability to transpose in the mouse germ line. First, the levels of activity of several Tc1͞mariner elements in mammalian cells were compared; the reconstructed fish transposon Sleeping Beauty (SB) was found to be an order of magnitude more efficient than the other tested transposons. SB then was introduced into the mouse germ line as a two-component system: one transgene for the expression of the transposase in the male germ line and a second transgene carrying a modified transposon. In 20% of the progeny of double transgenic male mice the transposon had jumped from the original chromosomal position into another locus. Analysis of the integration sites shows that these jumps indeed occurred through the action of SB transposase, and that SB has a strong preference for intrachromosomal transposition. Analysis of the excision sites suggests that double-strand breaks in haploid spermatids are repaired via nonhomologous end joining. The SB system may be a powerful tool for transposon mutagenesis of the mouse germ line.T ransposon tagging is a valuable tool for functional genomics in model organisms such as Drosophila (P-element) and Caenorhabditis elegans (Tc1 transposon). In the mouse germ line, however, efficient in vivo transposon tagging has not yet been achieved.Unlike P-element, members of the Tc1͞mariner superfamily of transposable elements do not require host-specific factors for their activity in vitro (1, 2). Indeed, several of these cut-and-paste transposons were shown to be active in a wide variety of species other than their original hosts, including bacteria [Himar1 (3)], malaria mosquito [Minos (4)], zebrafish [Tc3 (5) and Mos1 (6)], chicken [Mos1 (7)], and human cells [Tc1 (8), Sleeping Beauty (SB) (9), Himar1 (10), and Minos (11)]. Although the reconstructed fish transposon SB has been shown be able to jump in mouse embryonic stem (ES) cells (12), and more recently in mouse somatic tissue (13), germ-line transposition-an important and powerful tool-has yet to be demonstrated.To assess the utility of Tc1͞mariner elements for transposon tagging in the mouse germ line, we first compared Tc1, Tc3, Himar1, Mos1, and SB in an in vitro mammalian cell culture assay and determined that SB is most efficient. Based on these results, we introduced SB in the mouse germ line. We show that SB is able to jump with a high efficiency in the mouse germ line, demonstrating its potential utility for genetic applications. Materials and MethodsPlasmids. PCR fragments of the ORFs encoding the transposase proteins of Tc1, Tc3, Himar1, and Mos1 were cloned into the Klenow-treated, 3.8-kb NotI fragment of pCMV␤ (CLON-TECH), resulting in, respectively, pRP1341, pRP1342, pRP1389, and pRP1353. The template plasmids were, respectively, pRP470 (14), pRP716 (15), pMar27fH (2), and pMos1 (16). The mu...

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“…1D, Kaufman & Rio 1992;van Luenen et al 1994;Craig 1996;Kleckner et al 1996;Vos et al 1996). Double strand cleavage at both ends is obligatory prior to target DNA capture and strand transfer for Tn10 .…”

Section: Classical Transposition Reactionsmentioning

confidence: 99%

Polynucleotidyl transfer reactions in site‐specific DNA recombination

1997

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Site-specific DNA rearrangement reactions are widespread among organisms. They are used, for example, by vertebrates to boost immune response diversity, and in turn by parasitic organisms to evade the host immune system by surface antigen switching. Parasitic genetic elements ubiquitous to most organisms invade new host genomic sites by a variety of types of site-specific recombination. Polynucleotidyl transfer reactions are central to these DNA recombination reactions. The recombinase of each reaction system that 'catalyses' such chemical reactions at specific DNA sites are apparently designed to accomplish unique DNA geometrical specificity, or delicate control over the extent or direction of the reaction, with the sacrifice of protein turnover. Here we discuss our current understanding of several issues that relate to the polynucleotidyl transfer steps in several of the better studied site-specific recombination reactions.

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Transposase is the only nematode protein required for in vitro transposition of Tc1.

Cited by 176 publications

References 37 publications

The Tol1 element of the medaka fish, a member of the hAT transposable element family, jumps in Caenorhabditis elegans

The Tol1 element of the medaka fish, a member of the hAT transposable element family, jumps in Caenorhabditis elegans

Regulated transposition of a fish transposon in the mouse germ line

Polynucleotidyl transfer reactions in site‐specific DNA recombination

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