Gpr63 is a modifier of microcephaly in Ttc21b mouse mutants


Autoři: John Snedeker aff001;  William J. Gibbons, Jr. aff001;  David F. Paulding aff001;  Zakia Abdelhamed aff001;  Daniel R. Prows aff001;  Rolf W. Stottmann aff001
Působiště autorů: Division of Human Genetics, Cincinnati Children’s Hospital Medical Center, Cincinnati, Ohio, United States of America aff001;  Neuroscience Graduate Program, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States of America aff002;  Department of Anatomy and Embryology, Faculty of Medicine (Girl’s Section), Al-Azhar University, Cairo, Egypt aff003;  Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States of America aff004;  Division of Developmental Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, Ohio, United States of America aff005;  Shriner’s Hospital for Children - Cincinnati, Cincinnati, Ohio, United States of America aff006
Vyšlo v časopise: Gpr63 is a modifier of microcephaly in Ttc21b mouse mutants. PLoS Genet 15(11): e32767. doi:10.1371/journal.pgen.1008467
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008467

Souhrn

The primary cilium is a signaling center critical for proper embryonic development. Previous studies have demonstrated that mice lacking Ttc21b have impaired retrograde trafficking within the cilium and multiple organogenesis phenotypes, including microcephaly. Interestingly, the severity of the microcephaly in Ttc21baln/aln homozygous null mutants is considerably affected by the genetic background and mutants on an FVB/NJ (FVB) background develop a forebrain significantly smaller than mutants on a C57BL/6J (B6) background. We performed a Quantitative Trait Locus (QTL) analysis to identify potential genetic modifiers and identified two regions linked to differential forebrain size: modifier of alien QTL1 (Moaq1) on chromosome 4 at 27.8 Mb and Moaq2 on chromosome 6 at 93.6 Mb. These QTLs were validated by constructing congenic strains. Further analysis of Moaq1 identified an orphan G-protein coupled receptor (GPCR), Gpr63, as a candidate gene. We identified a SNP that is polymorphic between the FVB and B6 strains in Gpr63 and creates a missense mutation predicted to be deleterious in the FVB protein. We used CRISPR-Cas9 genome editing to create two lines of FVB congenic mice: one with the B6 sequence of Gpr63 and the other with a deletion allele leading to a truncation of the GPR63 C-terminal tail. We then demonstrated that Gpr63 can localize to the cilium in vitro. These alleles affect ciliary localization of GPR63 in vitro and genetically interact with Ttc21baln/aln as Gpr63;Ttc21b double mutants show unique phenotypes including spina bifida aperta and earlier embryonic lethality. This validated Gpr63 as a modifier of multiple Ttc21b neural phenotypes and strongly supports Gpr63 as a causal gene (i.e., a quantitative trait gene, QTG) within the Moaq1 QTL.

Klíčová slova:

Alleles – Cilia – Embryos – Mammalian genomics – Molecular genetics – Phenotypes – Quantitative trait loci – Sequence motif analysis


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Štítky
Genetika Reprodukční medicína

Článek vyšel v časopise

PLOS Genetics


2019 Číslo 11

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