What is the G542X mutation?

Mice homozygous for the G542X mutation have reduced Cftr expression and absence of CFTR function in the airway and intestine. These mice display typical cystic fibrosis manifestations such as poor growth and reduced survival due to intestinal obstruction.

What is the consequence of the F508 mutation?

The delta F508 mutation causes a diminution in the amount of beta-stranded structure and a concomitant increase in the amount of random coil structure present, indicating that either the mutant peptide has a different native structure or that the conformational equilibrium is shifted toward a more disordered form.

What class of mutation is Delta F508?

Class II includes ΔF508 for which the mutant protein is made in its entirety but fails to fold properly inside the cell, resulting in the absence of mature CFTR in the cellular membrane to perform its specific chloride ion conductance function.

Is cystic fibrosis a missense mutation?

General cystic fibrosis mutations are usually missense mutations affecting two specific protein domains and associated with a specific RFLP marker haplotype.

Can a G542X mouse model be used for CFTR?

Importantly, we demonstrate that pharmacological readthrough of the G542X nonsense mutation in this model allows production of functional CFTR. The G542X mouse will be a valuable model for the examination of nonsense mutation therapies for CF and other genetic diseases caused by nonsense mutations.

How are G542X Mice affected by cystic fibrosis?

Mice homozygous for the G542X mutation have reduced Cftr expression and absence of CFTR function in the airway and intestine. These mice display typical cystic fibrosis manifestations such as poor growth and reduced survival due to intestinal obstruction.

What kind of gRNA is used for G542X?

One gRNA (25 ng/ul; PNABio), a 120 bp single stranded oligonucleotide (ssODN) containing the G542X mutation (25 ng/ul; IDT) centered on the cut site and either Cas9 mRNA (25–50 ng/ul; PNABio) or Cas9 protein (25 ng/ul; PNABio) were injected in the pronucleus of C57BL/6 one-cell embryos.

How are nonsense mutations related to CFTR function?

Nonsense mutations are present in 10% of patients with CF, produce a premature termination codon in CFTR mRNA causing early termination of translation, and lead to lack of CFTR function.