Progress toward generating a ferret model of cystic fibrosis by somatic cell nuclear transfer

Abstract

Mammalian cloning by nuclear transfer from somatic cells has created new opportunities to generate animal models of genetic diseases in species other than mice. Although genetic mouse models play a critical role in basic and applied research for numerous diseases, often mouse models do not adequately reproduce the human disease phenotype. Cystic fibrosis (CF) is one such disease. Targeted ablation of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in mice does not adequately replicate spontaneous bacterial infections observed in the human CF lung. Hence, several laboratories are pursuing alternative animal models of CF in larger species such as the pig, sheep, rabbits, and ferrets. Our laboratory has focused on developing the ferret as a CF animal model. Over the past few years, we have investigated several experimental parameters required for gene targeting and nuclear transfer (NT) cloning in the ferret using somatic cells. In this review, we will discuss our progress and the hurdles to NT cloning and gene-targeting that accompany efforts to generate animal models of genetic diseases in species such as the ferret.

Optimizations of In Vitro Mucus and Cell Culture Models to Better Predict In Vivo Gene Transfer in Pathological Lung Respiratory Airways: Cystic Fibrosis as an Example

“…CFTR-knockout ferrets can be used as a potential model due to a lung structure close to that of humans. They exhibit altered chloride transport, leading to mucus hypersecretion and lung infections [ 32 , 33 ]. CF pigs also represent a potential model, as they present pulmonary manifestations similar to those found in human CF lung (infection, inflammation, airway remodeling, and mucus hypersecretion).…”

Comparative Biology of Cystic Fibrosis Animal Models

“…Several parameters influence the choice of alternative species to model CF including (1) the types of cells in the airway in comparison to human (2), the distribution of submucosal glands which are thought to play an important role in CF airway disease (3), conservation of CFTR structure and function (4), the composite of alternative chloride channels in the airway, and (5) the reproductive parameters of the species which will make it feasible to rapidly perform research studies. Generation of larger CF animal models such as pig (35), ferret (36), and sheep (37) has been considered, but the technology to manipulate the genomes of these animals has lagged behind until recently. Recently, methods for generating both pig and ferret CF models have been developed using recombinant adeno-associated virus (rAAV)-mediated gene targeting of exon 10 (6, 7).…”

Ferret acute lung injury model induced by repeated nebulized lipopolysaccharide administration

“…One of the main advantages of a ferret lung injury model is its lung biology and physiology that is similar to humans. Ferrets have portions of the upper and lower respiratory tracts, as well the number of generations of terminal bronchioles, and expression of relevant receptors and mutations (such as influenza receptor and CFTR mutation) that are similar to humans, resulting in similar disease pathogenesis (Johnson‐Delaney & Orosz, 2011 ; Leigh et al, 1986 ; Li & Engelhardt, 2003 ; Munoz‐Fontela et al, 2020 ; Oldham et al, 1990 ; Plopper et al, 1980 ). For this reason, ferrets have been used extensively as models for human respiratory diseases such as cystic fibrosis and viral infections like influenza and SARS‐COV‐2.…”