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- W2130506816 abstract "The clustered, regularly interspaced palindromic repeat (CRISPR) system is capable of genome-specific targeting utilizing the CRISPR associated 9 (Cas9) endoribonuclease and effector targeting by a single guide RNA (sgRNA). We show that fusion of the transcriptional activating viral protein, VP64, to an enzymatically dead form of Cas9 (dCas9) is capable of enhancing gene-targeted transcriptional activity. Using a luciferase reporter assay we show that a N-terminal fusion of VP64 to dCas9 (dCas9.nVP64) results in higher transcriptional activity compared to the C-terminal or flanked VP64 fusion to dCas9. Additionally, we find that there are optimal targets within the murine Tissue Inhibitor of Metalloproteinase 2 (TIMP2) promoter that results in maximal transcriptional activation and that multiple sgRNAs expressed simultaneously results in a synergistic enhancement of promoter activity. Finally, using a genome-wide bioinformatics approach we identified a number of potential off target sites for a given sgRNA-target within the promoter of other genes. Selective activation of TIMP2 with the CRISPR-Cas9 system opens up a strategy for treating diseases associated with over activity of the Matrix Metalloproteinases. Grant Funding Source: Supported by the Training Program in Translational Cardiovascular Science, 5 T32 HL 7936-12" @default.
- W2130506816 created "2016-06-24" @default.
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- W2130506816 date "2014-04-01" @default.
- W2130506816 modified "2023-09-25" @default.
- W2130506816 title "RNA‐guided transcriptional activation of TIMP2 utilizing CRISPR:Cas9 (1148.13)" @default.
- W2130506816 doi "https://doi.org/10.1096/fasebj.28.1_supplement.1148.13" @default.
- W2130506816 hasPublicationYear "2014" @default.
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