Target site selection by the mariner-like element, Mos1

Crénès, Gwénaëlle; Moundras, Corinne; Demattei, Marie-Véronique; Bigot, Yves; Petit, Agnès; Renault, Sylvie

doi:10.1007/s10709-009-9387-6

Cited by 20 publications

(17 citation statements)

References 39 publications

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“…We confirmed previous observations 31,32 that Mos1 shows no target site selection in vitro except for integration at TA nucleotides. Our work shows that Mboumar-9 transposase behaves similarly; no integration site specificity was noticed for Mboumar-9 transposition in vitro .…”

Section: Discussionsupporting

confidence: 92%

Biochemical Characterization and Comparison of Two Closely Related Active mariner Transposases

et al. 2014

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Most DNA transposons move from one genomic location to another by a cut-and-paste mechanism and are useful tools for genomic manipulations. Short inverted repeat (IR) DNA sequences marking each end of the transposon are recognized by a DNA transposase (encoded by the transposon itself). This enzyme cleaves the transposon ends and integrates them at a new genomic location. We report here a comparison of the biophysical and biochemical properties of two closely related and active mariner/Tc1 family DNA transposases: Mboumar-9 and Mos1. We compared the in vitro cleavage activities of the enzymes on their own IR sequences, as well as cross-recognition of their inverted repeat sequences. We found that, like Mos1, untagged recombinant Mboumar-9 transposase is a dimer and forms a stable complex with inverted repeat DNA in the presence of Mg2+ ions. Mboumar-9 transposase cleaves its inverted repeat DNA in the manner observed for Mos1 transposase. There was minimal cross-recognition of IR sequences between Mos1 and Mboumar-9 transposases, despite these enzymes having 68% identical amino acid sequences. Transposases sharing common biophysical and biochemical properties, but retaining recognition specificity toward their own IR, are a promising platform for the design of chimeric transposases with predicted and improved sequence recognition.

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Section: Discussionsupporting

confidence: 92%

Biochemical Characterization and Comparison of Two Closely Related Active mariner Transposases

et al. 2014

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“…This plasmid was unable to confer resistance to Escherichia coli cells in tetracycline concentrations over 10 µg/ml. Upon transposition, MOS1 excises the pseudo- Mos1 from one pBC-3T3 molecule, and then triggers its reinsertion within the cat gene of another pBC-3T3 molecule [28]. Transposition events are revealed by promoter tagging, the tetracycline resistance being activated through the cat gene promoter.…”

Section: Resultsmentioning

confidence: 99%

Natural Stilbenoids Isolated from Grapevine Exhibiting Inhibitory Effects against HIV-1 Integrase and Eukaryote MOS1 Transposase In Vitro Activities

et al. 2013

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Polynucleotidyl transferases are enzymes involved in several DNA mobility mechanisms in prokaryotes and eukaryotes. Some of them such as retroviral integrases are crucial for pathogenous processes and are therefore good candidates for therapeutic approaches. To identify new therapeutic compounds and new tools for investigating the common functional features of these proteins, we addressed the inhibition properties of natural stilbenoids deriving from resveratrol on two models: the HIV-1 integrase and the eukaryote MOS-1 transposase. Two resveratrol dimers, leachianol F and G, were isolated for the first time in Vitis along with fourteen known stilbenoids: E-resveratrol, E-piceid, E-pterostilbene, E-piceatannol, (+)-E-ε-viniferin, E-ε-viniferinglucoside, E-scirpusin A, quadragularin A, ampelopsin A, pallidol, E-miyabenol C, E-vitisin B, hopeaphenol, and isohopeaphenol and were purified from stalks of Vitis vinifera (Vitaceae), and moracin M from stem bark of Milliciaexelsa (Moraceae). These compounds were tested in in vitro and in vivo assays reproducing the activity of both enzymes. Several molecules presented significant inhibition on both systems. Some of the molecules were found to be active against both proteins while others were specific for one of the two models. Comparison of the differential effects of the molecules suggested that the compounds could target specific intermediate nucleocomplexes of the reactions. Additionally E-pterostilbene was found active on the early lentiviral replication steps in lentiviruses transduced cells. Consequently, in addition to representing new original lead compounds for further modelling of new active agents against HIV-1 integrase, these molecules could be good tools for identifying such reaction intermediates in DNA mobility processes.

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“…Electroporation is difficult to automate; thus protocols, which do not require electroporation offer the potential for automated high-throughput generation of insertion libraries. Mariner transposases can integrate transposons in any TA dinucleotide present in the genome (36,48), providing an advantage for full genome coverage.…”

Section: Discussionmentioning

confidence: 99%

Use of mariner transposases for one-step delivery and integration of DNA in prokaryotes and eukaryotes by transfection

Trubitsyna

Michlewski

Finnegan

et al. 2017

Nucleic Acids Research

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Delivery of DNA to cells and its subsequent integration into the host genome is a fundamental task in molecular biology, biotechnology and gene therapy. Here we describe an IP-free one-step method that enables stable genome integration into either prokaryotic or eukaryotic cells. A synthetic mariner transposon is generated by flanking a DNA sequence with short inverted repeats. When purified recombinant Mos1 or Mboumar-9 transposase is co-transfected with transposon-containing plasmid DNA, it penetrates prokaryotic or eukaryotic cells and integrates the target DNA into the genome. In vivo integrations by purified transposase can be achieved by electroporation, chemical transfection or Lipofection of the transposase:DNA mixture, in contrast to other published transposon-based protocols which require electroporation or microinjection. As in other transposome systems, no helper plasmids are required since transposases are not expressed inside the host cells, thus leading to generation of stable cell lines. Since it does not require electroporation or microinjection, this tool has the potential to be applied for automated high-throughput creation of libraries of random integrants for purposes including gene knock-out libraries, screening for optimal integration positions or safe genome locations in different organisms, selection of the highest production of valuable compounds for biotechnology, and sequencing.

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Target site selection by the mariner-like element, Mos1

Cited by 20 publications

References 39 publications

Biochemical Characterization and Comparison of Two Closely Related Active mariner Transposases

Biochemical Characterization and Comparison of Two Closely Related Active mariner Transposases

Natural Stilbenoids Isolated from Grapevine Exhibiting Inhibitory Effects against HIV-1 Integrase and Eukaryote MOS1 Transposase In Vitro Activities

Use of mariner transposases for one-step delivery and integration of DNA in prokaryotes and eukaryotes by transfection

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