ArticleMolecular therapy : the journal of the American Society of Gene Therapy2009
Mutational derivatives of PhiC31 integrase with increased efficiency and specificity.
Article in Molecular therapy : the journal of the American Society of Gene Therapy, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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Who cites it
41 citing papers in PubMed, 68 citations in OpenAlex.
- Article
- Site-specific DNA insertion into the human genome with engineered recombinases.Nature biotechnology · 2025Article
- Directed evolution of hyperactive integrases for site specific insertion of transgenes.Nucleic acids research · 2024Article
- Directed evolution of hyperactive integrases for site specific insertion of transgenes.bioRxiv : the preprint server for biology · 2024Article
- A novel approach for T7 bacteriophage genome integration of exogenous DNA.Journal of biological engineering · 2020Article
- Engineering altered protein-DNA recognition specificity.Nucleic acids research · 2018Review
- Temporal self-regulation of transposition through host-independent transposase rodlet formation.Nucleic acids research · 2017Article
- Enhanced heterologous protein productivity by genome reduction in Lactococcus lactis NZ9000.Microbial cell factories · 2017Article
- Ultrasound-targeted hepatic delivery of factor IX in hemophiliac mice.Gene therapy · 2016Article
- Exogenous enzymes upgrade transgenesis and genetic engineering of farm animals.Cellular and molecular life sciences : CMLS · 2015Review
- Article
- Visualizing genome and systems biology: technologies, tools, implementation techniques and trends, past, present and future.GigaScience · 2015Review
- Highly modular bow-tie gene circuits with programmable dynamic behaviour.Nature communications · 2014Article
- Serine integrase chimeras with activity in E. coli and HeLa cells.Biology open · 2014Article
- Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase.BMC biotechnology · 2014Article
- PhiC31 integrase induces efficient site-specific recombination in the Capra hircus genome.DNA and cell biology · 2014Article
- Enhancing the specificity of recombinase-mediated genome engineering through dimer interface redesign.Journal of the American Chemical Society · 2014Article
- Expanding the scope of site-specific recombinases for genetic and metabolic engineering.Biotechnology and bioengineering · 2014Review
- Attachment site recognition and regulation of directionality by the serine integrases.Nucleic acids research · 2013Article
- Article
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
Abstract
phiC31 integrase is a sequence-specific phage recombinase that can recombine two short DNA sequences called attB and attP. The enzyme can also promote genomic integration of plasmids carrying attB into native mammalian sequences having partial identity to attP. To increase the efficiency of integration, we mutated the phiC31 integrase gene and screened the mutants in human cells in an assay for higher recombination frequency between attB and attP. We report in this article the isolation of a mutant, P2 that has twice the chromosomal integration frequency of wild-type phiC31 integrase, at both a preintegrated chromosomal attP site and at endogenous pseudo attP sequences in cultured human cells. In mouse liver, P2-mediated integration provided therapeutic long-term levels of human factor IX that were double those generated by wild-type phiC31 integrase. We also describe an additional mutant, P3 that combines the mutations of P2 with further changes and possesses an elevated specificity for integration at a chromosomally placed attP site in human cells. Forty-four percent of colonies carrying integration events mediated by P3 have integration at the placed attP site. These mutant integrases are useful for gene therapy and genome modification, and they demonstrate the feasibility of engineering phiC31 integrase toward more desirable properties.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.