TY - JOUR
TI - A BAC Transgene Expressing Human CFTR under Control of Its Regulatory Elements Rescues Cftr Knockout Mice
AU - Gawenis, L.R.
AU - Hodges, C.A.
AU - McHugh, D.R.
AU - Valerio, D.M.
AU - Miron, A.
AU - Cotton, C.U.
AU - Liu, J.
AU - Walker, N.M.
AU - Strubberg, A.M.
AU - Gillen, A.E.
AU - Mutolo, M.J.
AU - Kotzamanis, G.
AU - Bosch, J.
AU - Harris, A.
AU - Drumm, M.L.
AU - Clarke, L.L.
JO - Scientific Reports
PY - 2019
VL - 9
TODO - 1
SP - null
PB - Nature Publishing Group
SN - 2045-2322
TODO - 10.1038/s41598-019-48105-4
TODO - CFTR protein, human;  cystic fibrosis transmembrane conductance regulator, animal;  bacterial artificial chromosome;  C57BL mouse;  exocytosis;  gene editing;  genetics;  human;  knockout mouse;  male;  mouse;  regulatory sequence;  transgene;  transgenic mouse, Animals;  Chromosomes, Artificial, Bacterial;  Cystic Fibrosis Transmembrane Conductance Regulator;  Exocytosis;  Gene Editing;  Humans;  Male;  Mice;  Mice, Inbred C57BL;  Mice, Knockout;  Mice, Transgenic;  Regulatory Sequences, Nucleic Acid;  Transgenes
TODO - Small-molecule modulators of cystic fibrosis transmembrane conductance regulator (CFTR) biology show promise in the treatment of cystic fibrosis (CF). A Cftr knockout (Cftr KO) mouse expressing mutants of human CFTR would advance in vivo testing of new modulators. A bacterial artificial chromosome (BAC) carrying the complete hCFTR gene including regulatory elements within 40.1 kb of DNA 5′ and 25 kb of DNA 3′ to the gene was used to generate founder mice expressing hCFTR. Whole genome sequencing indicated a single integration site on mouse chromosome 8 (8qB2) with ~6 gene copies. hCFTR+ offspring were bred to murine Cftr KO mice, producing hCFTR+/mCftr− (H+/m−) mice, which had normal survival, growth and goblet cell function as compared to wild-type (WT) mice. Expression studies showed hCFTR protein and transcripts in tissues typically expressing mCftr. Functionally, nasal potential difference and large intestinal short-circuit (Isc) responses to cAMP stimulation were similar in magnitude to WT mice, whereas small intestinal cAMP ΔIsc responses were reduced. A BAC transgenic mouse with functional hCFTR under control of its regulatory elements has been developed to enable the generation of mouse models of hCFTR mutations by gene editing for in vivo testing of new CF therapies. © 2019, The Author(s).
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