ArticleBMC biotechnology2010
Comparative analysis of right element mutant lox sites on recombination efficiency in embryonic stem cells.
Article in BMC biotechnology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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16 citing papers in PubMed, 41 citations in OpenAlex.
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- The lncRNA Caren antagonizes heart failure by inactivating DNA damage response and activating mitochondrial biogenesis.Nature communications · 2021Article
- Understanding and exploiting the fatty acid desaturation system in Rhodotorula toruloides.Biotechnology for biofuels · 2021Article
- Lox'd in translation: contradictions in the nomenclature surrounding common lox-site mutants and their implications in experiments.Microbiology (Reading, England) · 2021Article
- iDamage: a method to integrate modified DNA into the yeast genome.Nucleic acids research · 2019Article
- Testing the Cre-mediated genetic switch for the generation of conditional knock-in mice.PloS one · 2019Article
- A photoactivatable Cre-loxP recombination system for optogenetic genome engineering.Nature chemical biology · 2016Article
- Gene stacking by recombinases.Plant biotechnology journal · 2016Review
- The kick-in system: a novel rapid knock-in strategy.PloS one · 2014Article
- Generation and analysis of serine protease inhibitor kazal type 3-cre driver mice.Experimental animals · 2014Article
- Improved tools for the Brainbow toolbox.Nature methods · 2013Article
- Ectopic expression of Ptf1a induces spinal defects, urogenital defects, and anorectal malformations in Danforth's short tail mice.PLoS genetics · 2013Article
- RAD51 mutants cause replication defects and chromosomal instability.Molecular and cellular biology · 2012Article
- Recombinase technology: applications and possibilities.Plant cell reports · 2011Review
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4 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
backgroundCre-mediated site-specific integrative recombination in mouse embryonic stem (ES) cells is a useful tool for genome engineering, allowing precise and repeated site-specific integration. To promote the integrative reaction, a left element/right element (LE/RE) mutant strategy using a pair of lox sites with mutations in the LE or RE of the lox sequence has previously been developed. Recombination between LE and RE mutant lox produces a wild-type loxP site as well as an LE+RE double mutant lox site, which has mutations in both sides and less affinity to Cre, resulting in stable integration. We previously demonstrated successful integrative recombination using lox71 (an LE mutant) and lox66 (an RE mutant) in ES cells. Recently, other LE/RE mutant lox sites showing higher recombination efficiency in Escherichia coli have been reported. However, their recombination efficiency in mammalian cells remains to be analyzed.
resultsUsing ES cells, we compared six RE mutant lox sites, focusing on their recombination efficiency with lox71. All of the RE mutant lox sites showed similar recombination efficiency. We then analyzed the stability of the recombined product, i.e., the LE+RE double mutant lox site, under continuous and strong Cre activity in ES cells. Two RE mutants, loxJTZ17 and loxKR3, produced more stable LE+RE double mutant lox than did the lox66/71 double mutant.
conclusionThe two mutant RE lox sites, loxJTZ17 and loxKR3, are more suitable than lox66 for Cre-mediated integration or inversion in ES cells.
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