CRISPR/Cas9 interrogation of the mouse Pcdhg gene cluster reveals a crucial isoform-specific role for Pcdhgc4.

Autor: Garrett AM; Department of Pharmacology and Department of Ophthalmology, Visual, and Anatomical Sciences, Wayne State University, Detroit, Michigan, United States of America.; The Jackson Laboratory, Bar Harbor, Maine, United States of America., Bosch PJ; Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of America., Steffen DM; Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of America., Fuller LC; Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of America., Marcucci CG; Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of America., Koch AA; Department of Pharmacology and Department of Ophthalmology, Visual, and Anatomical Sciences, Wayne State University, Detroit, Michigan, United States of America., Bais P; The Jackson Laboratory, Bar Harbor, Maine, United States of America., Weiner JA; Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of America., Burgess RW; The Jackson Laboratory, Bar Harbor, Maine, United States of America.
Jazyk: angličtina
Zdroj: PLoS genetics [PLoS Genet] 2019 Dec 26; Vol. 15 (12), pp. e1008554. Date of Electronic Publication: 2019 Dec 26 (Print Publication: 2019).
DOI: 10.1371/journal.pgen.1008554
Abstrakt: The mammalian Pcdhg gene cluster encodes a family of 22 cell adhesion molecules, the gamma-Protocadherins (γ-Pcdhs), critical for neuronal survival and neural circuit formation. The extent to which isoform diversity-a γ-Pcdh hallmark-is required for their functions remains unclear. We used a CRISPR/Cas9 approach to reduce isoform diversity, targeting each Pcdhg variable exon with pooled sgRNAs to generate an allelic series of 26 mouse lines with 1 to 21 isoforms disrupted via discrete indels at guide sites and/or larger deletions/rearrangements. Analysis of 5 mutant lines indicates that postnatal viability and neuronal survival do not require isoform diversity. Surprisingly, given reports that it might not independently engage in trans-interactions, we find that γC4, encoded by Pcdhgc4, is the only critical isoform. Because the human orthologue is the only PCDHG gene constrained in humans, our results indicate a conserved γC4 function that likely involves distinct molecular mechanisms.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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