Projects per year
Abstract / Description of output
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.
Original language | English |
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Article number | e89 |
Number of pages | 10 |
Journal | Nucleic Acids Research |
Volume | 45 |
Issue number | 10 |
Early online date | 16 Feb 2017 |
DOIs | |
Publication status | Published - 2 Jun 2017 |
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Dive into the research topics of 'Use of mariner transposases for one-step delivery and integration of DNA in prokaryotes and eukaryotes by transfection'. Together they form a unique fingerprint.Projects
- 3 Finished
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SynthSys-Mammalian: Edinburgh Mammalian Synthetic Biology Research Centre
14/11/14 → 31/03/22
Project: Research
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Regulation of miRNA Biogenesis and function in humans
Michlewski, G.
1/01/11 → 31/12/15
Project: Research
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Profiles
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Alistair Elfick
- School of Engineering - Personal Chair of Synthetic Biological Engineering
- Centre for Engineering Biology
Person: Academic: Research Active
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Chris French
- School of Biological Sciences - Personal Chair of Microbial Biotechnology
- Centre for Engineering Biology
Person: Academic: Research Active