Disruption of the CFTR Gene Produces a Model of Cystic Fibrosis in Newborn Pigs

Abstract

SUMMARY Almost two decades after identification of the CFTR gene, we lack answers to many questions about the pathogenesis of cystic fibrosis (CF), and it remains a lethal disease. Mice with a disrupted CFTR gene have greatly facilitated CF studies, but they fail to develop the characteristic pancreatic, lung, intestinal, liver, and other CF manifestations. Therefore, we produced pigs with a targeted disruption of both CFTR alleles. These animals exhibited defective chloride transport. They also developed meconium ileus, exocrine pancreatic destruction, and focal biliary cirrhosis, replicating abnormalities seen in newborn patients with CF. This swine model may provide opportunities to address persistent questions about CF pathogenesis and accelerate discovery of treatments and preventions.

Germline transgenesis in pigs by cytoplasmic microinjection of Sleeping Beauty transposons

“…The procedure includes gene transfer into primary somatic cells, screening of cells carrying the desired genetic modification, introduction of a genetically modified cell into an enucleated porcine oocyte and activation of the reconstructed embryo to initiate cleavage divisions 3,7,8 . The transfer of >100 reconstructed embryos to one surrogate animal is required to establish a pregnancy in the pig 7,8 and success rates of only 1-3 % per reconstructed embryos have been achieved 7,8 . Furthermore, SCNT protocols are associated with a high workload, and partial genome reprogramming during SCNT commonly results in high rates of abortion and neonatal deaths.…”

“…In recent years, improved methods for transgenesis have made this model even more valuable 5 . Humanized pig models include those for retinitis pigmentosa 6 , cystic fibrosis 7 , Alzheimer´s disease 8 , Huntington´s disease 9 , familial adenomatous polyposis 10 , and immunodeficiency 11 . However, transgenesis in the pig, most commonly achieved by pronuclear DNA injection (PNI) or by somatic cell nuclear transfer (SCNT), is an inefficient and expensive process, hampered by poor predictability of levels and patterns of transgene expression 2,3,[12][13][14] .…”

Antibody Repertoire Development in Fetal and Neonatal Piglets. XX. B Cell Lymphogenesis Is Absent in the Ileal Peyer’s Patches, Their Repertoire Development Is Antigen Dependent, and They Are Not Required for B Cell Maintenance

“…Addressing the role of the IPP has implications beyond comparative and veterinary immunology because swine are used as models for human disease in cases where mouse models fail (50)(51)(52), in studies on xenotransplantation (53), as a model for immuno-ontogeny (41), and in the generation of B cell knockout pigs as the first step toward preparing humanized Abs in a large xenogeneic species (54,55).…”

Genetically manipulated mouse models of lung disease: potential and pitfalls

“…The ability to routinely perform genetic modification in larger mammals will expand our capabilities of carrying out clinically relevant research. Interestingly, Rogers et al (128,129) used this rationale to generate pigs with targeted disruption of the cystic fibrosis transmembrane receptor gene (CFTR) and found that CFTR-deficient (CFTR Ϫ/Ϫ ) pigs recapitulated human cystic fibrosis far more accurately than did CFTR Ϫ/Ϫ mice. More recently, Tong et al (154) demonstrated the feasibility of gene targeting in rats through homologous recombination in pluripotent rat ES cells, thus expanding dramatically the future repertoire of possible experiments using rat-based systems.…”