ArticleNucleic acids research2013
rDNA-directed integration by an HIV-1 integrase--I-PpoI fusion protein.
Article in Nucleic acids research, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Biology and utilization of R2 retrotransposons.RNA biology · 2025Review
- Advances in HIV Gene Therapy.International journal of molecular sciences · 2024Review
- Article
- Article
- Strategies for Targeting Retroviral Integration for Safer Gene Therapy: Advances and Challenges.Frontiers in molecular biosciences · 2021Review
- Efficient Nuclease-Directed Integration of Lentivirus Vectors into the Human Ribosomal DNA Locus.Molecular therapy : the journal of the American Society of Gene Therapy · 2020Article
- Targeted gene knockin in zebrafish using the 28S rDNA-specific non-LTR-retrotransposon R2Ol.Mobile DNA · 2019Article
- Paired CRISPR/Cas9 Nickases Mediate Efficient Site-Specific Integration ofInternational journal of molecular sciences · 2018Article
- Time-Restricted PiggyBac DNA Transposition by Transposase Protein Delivery Using Lentivirus-Derived Nanoparticles.Molecular therapy. Nucleic acids · 2018Article
- Retroviral vector interactions with hematopoietic cells.Current opinion in virology · 2016Review
- Retargeted Foamy Virus Vectors Integrate Less Frequently Near Proto-oncogenes.Scientific reports · 2016Article
- Nucleolar Reorganization Upon Site-Specific Double-Strand Break Induction.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2016Article
- Driving DNA transposition by lentiviral protein transduction.Mobile genetic elements · 2014Article
- Lentiviral protein transduction with genome-modifying HIV-1 integrase-I-PpoI fusion proteins: studies on specificity and cytotoxicity.BioMed research international · 2014Article
- Nonviral gene targeting at rDNA locus of human mesenchymal stem cells.BioMed research international · 2013Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Integrating viral vectors are efficient gene transfer tools, but their integration patterns have been associated with genotoxicity and oncogenicity. The recent development of highly specific designer nucleases has enabled target DNA modification and site-specific gene insertion at desired genomic loci. However, a lack of consensus exists regarding a perfect genomic safe harbour (GSH) that would allow transgenes to be stably and reliably expressed without adversely affecting endogenous gene structure and function. Ribosomal DNA (rDNA) has many advantages as a GSH, but efficient means to target integration to this locus are currently lacking. We tested whether lentivirus vector integration can be directed to rDNA by using fusion proteins consisting of the Human Immunodeficiency Virus 1 (HIV-1) integrase (IN) and the homing endonuclease I-PpoI, which has natural cleavage sites in the rDNA. A point mutation (N119A) was introduced into I-PpoI to abolish unwanted DNA cleavage by the endonuclease. The vector-incorporated IN-I-PpoIN119A fusion protein targeted integration into rDNA significantly more than unmodified lentivirus vectors, with an efficiency of 2.7%. Our findings show that IN-fusion proteins can be used to modify the integration pattern of lentivirus vectors, and to package site-specific DNA-recognizing proteins into vectors to obtain safer transgene integration.
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