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| Content Provider | PubMed Central |
|---|---|
| Author | Turkki, Vesa Diana, Schenkwein Timonen, Oskari Husso, Tiia Lesch, Hanna P. Seppo, Ylä-herttuala |
| Copyright Year | 2014 |
| Abstract | Rare-cutting endonucleases, such as the I-PpoI, can be used for the induction of double strand breaks (DSBs) in genome editing and targeted integration based on homologous recombination. For therapeutic approaches, the specificity and the pattern of off-target effects are of high importance in these techniques. For its applications, the endonuclease needs to be transported into the target cell nucleus, where the mechanism of transport may affect its function. Here, we have studied the lentiviral protein transduction of the integrase (IN)-PpoI fusion protein using the cis-packaging method. In genome-wide interaction studies, IN-fusion proteins were verified to bind their target sequence containing 28S ribosomal RNA (rRNA) genes with a 100-fold enrichment, despite the well-documented behavior of IN to be tethered into various genomic areas by host-cell factors. In addition, to estimate the applicability of the method, DSB-induced cytotoxic effects with different vector endonuclease configurations were studied in a panel of cells. Varying the amount and activity of endonuclease enabled the adjustment of ratio between the induced DSBs and transported DNA. In cell studies, certain cancerous cell lines were especially prone to DSBs in rRNA genes, which led us to test the protein transduction in a tumour environment in an in vivo study. In summary, the results highlight the potential of lentiviral vectors (LVVs) for the nuclear delivery of endonucleases. |
| Related Links | http://dx.doi.org/10.1155/2014/379340 |
| Starting Page | 379340 |
| File Format | |
| ISSN | 23146133 |
| e-ISSN | 23146141 |
| Journal | BioMed Research International |
| Volume Number | 2014 |
| Language | English |
| Publisher | Hindawi Publishing Corporation |
| Publisher Date | 2014-01-01 |
| Access Restriction | Open |
| Rights Holder | Hindawi Publishing Corporation |
| Subject Keyword | Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Immunology and Microbiology Medicine Biochemistry, Genetics and Molecular Biology |
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