title: Loss of Chd7 function in gene-trapped reporter mice is embryonic lethal and associated with severe defects in multiple developing tissues.
abstract: CHD7 is a novel chromodomain gene mutated in 60%-80% of humans with CHARGE syndrome, a multiple congenital anomaly condition characterized by ocular coloboma, heart defects, atresia of the choanae, retarded growth and development, genital hypoplasia, and characteristic ear abnormalities including deafness. Phenotypic features of CHARGE are highly variable and incompletely penetrant. To explore developmental roles of CHD7, we generated mice carrying the Chd7(Gt) allele from a Chd7-deficient, gene-trapped lacZ reporter ES cell line. RT-PCR of embryo RNA demonstrated significantly reduced levels of wild-type transcript in Chd7(Gt/Gt) embryos. Chd7(Gt/Gt) embryos survive only up to embryonic day 10.5 (E10.5). Chd7(Gt/+) male and female mice are viable, small, and exhibit variable degrees of head-bobbing and circling, consistent with vestibular dysfunction. Paint-filling of E16.5 heterozygous inner ears revealed defects of the semicircular canals. The pattern of beta-galactosidase activity in Chd7(Gt/+) embryos mimics Chd7 mRNA expression in wild-type embryos, confirming the fidelity of the lacZ reporter. We observed tissue-specific beta-galactosidase in the E12.5 and E14.5 Chd7(Gt/+) brain, pituitary, ear, heart, and craniofacial structures, indicating survival of Chd7(Gt/+) cells in CHARGE-relevant organs. These studies demonstrate the utility of Chd7(Gt) as a reporter-tagged loss-of-function allele for future studies exploring developmental mechanisms of Chd7 deficiency.
authors.affiliation: Department of Pediatrics, The University of Michigan Medical Center, Ann Arbor, Michigan 48109-0652, USA.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Hurd
authors.forename: Patrice L
authors.initials: PL
authors.lastname: Capers
authors.forename: Marsha N
authors.initials: MN
authors.lastname: Blauwkamp
authors.forename: Meredith E
authors.initials: ME
authors.lastname: Adams
authors.forename: Yehoash
authors.initials: Y
authors.lastname: Raphael
authors.forename: Heather K
authors.initials: HK
authors.lastname: Poucher
authors.forename: Donna M
authors.initials: DM
authors.lastname: Martin
mesh_terms: D000818:Animals; D004265:DNA Helicases / Q000235:genetics; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics*; D007758:Ear, Inner / Q000002:abnormalities; D053595:Embryonic Stem Cells / Q000502:physiology; D005804:Genes, Lethal; D017930:Genes, Reporter; D006580:Genetic Carrier Screening; D005838:Genotype; D006720:Homozygote; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C507367:Chd7 protein, mouse; D017931:DNA Primers; D004268:DNA-Binding Proteins; D004265:DNA Helicases