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CRISPR/Cas9 Editing of the p.F508del CFTR Mutation - Coggle Diagram
CRISPR/Cas9 Editing of the p.F508del CFTR Mutation
Disease
Cystic Fibrosis (CF)= caused by mutations in the CFTR gene.
mutation: p.F508del from deletion
Leads to defective chloride transport in epithelial cells.
Goal
Use CRISPR/Cas9 genome editing to correct the p.F508del mutation.
Improve precision and reduce off-target effects using Cas9 variants and sgRNAs.
Tools
SpCas9(1.1), SpCas9(HF4), SaCas9.
sgRNAs targeting the mutant CFTR locus
ssODN donor templates for precise repair
Dna repair
NHEJ (Non-Homologous End Joining) =error-prone, causes indels.
HDR (Homology-Directed Repair) = precise correction using donor template.
Results
Combination of SpCas9(HF4) + optimized sgRNA + ssODN produced highest HDR (~50%) in
CFTE29o- cells.
iPSCs showed higher NHEJ but lower HDR (~19–20%).
efficiency depends on cell type and CRISPR design.
CRISPR can correct p.F508del in vitro.
limiting for therapeutic application.