Nanoparticles that deliver triplex-forming peptide nucleic acid molecules correct F508del CFTR in airway epithelium.
Nicole Ali McNeerKavitha AnandalingamRachel J FieldsChristina CaputoSascha KopicAnisha GuptaElias QuijanoLee PolikoffYong KongRaman BahalJohn P GeibelPeter M GlazerW Mark SaltzmanMarie E EganPublished in: Nature communications (2015)
Cystic fibrosis (CF) is a lethal genetic disorder most commonly caused by the F508del mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. It is not readily amenable to gene therapy because of its systemic nature and challenges including in vivo gene delivery and transient gene expression. Here we use triplex-forming peptide nucleic acids and donor DNA in biodegradable polymer nanoparticles to correct F508del. We confirm modification with sequencing and a functional chloride efflux assay. In vitro correction of chloride efflux occurs in up to 25% of human cells. Deep-sequencing reveals negligible off-target effects in partially homologous sites. Intranasal delivery of nanoparticles in CF mice produces changes in the nasal epithelium potential difference assay, consistent with corrected CFTR function. Also, gene correction is detected in the nasal and lung tissue. This work represents facile genome engineering in vivo with oligonucleotides using a nanoparticle system to achieve clinically relevant levels of gene editing without off-target effects.
Keyphrases
- cystic fibrosis
- nucleic acid
- pseudomonas aeruginosa
- genome wide
- gene expression
- lung function
- gene therapy
- copy number
- high throughput
- dna methylation
- single cell
- drug delivery
- adipose tissue
- single molecule
- dna repair
- climate change
- transcription factor
- air pollution
- metal organic framework
- high fat diet induced
- highly efficient
- subarachnoid hemorrhage
- reduced graphene oxide
- blood brain barrier
- cell free
- drug induced