| 研究実績の概要 |
We changed nonsense codon (TAA) into sense codon (TGG) by using minidCas13X.1 and ADAR editor to the target site of the DMD transcript in a sequence-specific manner with the help of complementary guide RNAs. As a progress, we have made seven retroviral vectors (four guide RNAs vectors, two ADAR vectors with minidCas13X.1 and MS2 protein, one EGFP) and confirmed by sanger sequencing. To confirm the expression and retroviral packaging, we transduce the EGFP vectors into h2kmdx23 primary myoblast cells isolated from mdx23 mouse. The 70% cells have shown EGFP florescence according to microscopical observation. Which indicated that viral vector construction and packaging are good enough for next experiment. However, we have not yet checked the editing event in the cellular system.
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