Cystic fibrosis (CF) is caused by mutations in the gene encoding the transmembraneconductance regulator (CFTR) protein. Some CF patients are compound heterozygous or homozygousfor nonsense mutations in the CFTR gene. This implies the presence in the transcript of prematuretermination codons (PTCs) responsible for a truncated CFTR protein and a more severe formof the disease. Aminoglycoside and PTC124 derivatives have been used for the read-throughof PTCs to restore the full-length CFTR protein. However, in a precision medicine framework,the CRISPR/dCas13b-based molecular tool “REPAIRv2” (RNA Editing for Programmable A to IReplacement, version 2) could be a good alternative to restore the full-length CFTR protein. This RNAediting approach is based on the targeting of the deaminase domain of the hADAR2 enzyme fused tothe dCas13b protein to a specific adenosine to be edited to inosine in the mutant mRNA. Targetingspecificity is allowed by a guide RNA (gRNA) complementarily to the target region and recognizedby the dCas13b protein. Here, we used the REPAIRv2 platform to edit the UGA PTC to UGG indierent cell types, namely IB3-1 cells, HeLa, and FRT cells engineered to express H2BGFPopal andCFTRW1282X, respectively.
|Numero di pagine||16|
|Rivista||International Journal of Molecular Sciences|
|Stato di pubblicazione||Published - 2020|
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