2015 Fiscal Year Final Research Report
Genome editing tool for gene therapy in human hematopoietic stem cells
| Project/Area Number |
26670168
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Human genetics
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Keywords | ゲノム編集 / 遺伝子治療 / CRISPR / TALEN / 血液幹細胞 |
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| Outline of Final Research Achievements |
The Cas9 nucleases from the Streptococcus pyogenes, directed by a chimeric guide RNA (sgRNA) to any genomic locus followed by a 5’-NGG protospaceradjacent motif (PAM), are robust tools for inducing targeted double-stranded DNA breaks (DSBs) in human cells. The RNA-guided endonuclease (RGEN) was applied for human T cells, and then the system created recombinant cells. DSBs induce homology-directed repair (HDR). From the HDR pathway, site-directed insertion (knock-in, KI) was induced by provision of homologous donor construct and the RGENs significantly promote efficient its site-specific insertion in PSIP1 gene. We created to develop an efficient homozygous KI system via fluorescent gene transduction. We also developed an efficient TALEN-genome editing system to remove HIV proviral DNA.
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| Free Research Field |
ウイルス
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