Journal article
CRISPR/Cas9 interrogation of the mouse Pcdhg gene cluster reveals a crucial isoform-specific role for Pcdhgc4
PLoS genetics, Vol.15(12), pp.e1008554-e1008554
12/2019
DOI: 10.1371/journal.pgen.1008554
PMCID: PMC6957209
PMID: 31877124
Abstract
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.
Details
- Title: Subtitle
- CRISPR/Cas9 interrogation of the mouse Pcdhg gene cluster reveals a crucial isoform-specific role for Pcdhgc4
- Creators
- Andrew M Garrett - The Jackson Laboratory, Bar Harbor, Maine, United States of AmericaPeter J Bosch - Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of AmericaDavid M Steffen - Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of AmericaLeah C Fuller - Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of AmericaCharles G Marcucci - Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of AmericaAlexis A Koch - Department of Pharmacology and Department of Ophthalmology, Visual, and Anatomical Sciences, Wayne State University, Detroit, Michigan, United States of AmericaPreeti Bais - The Jackson Laboratory, Bar Harbor, Maine, United States of AmericaJoshua A Weiner - Department of Biology and Iowa Neuroscience Institute, University of Iowa, Iowa City, Iowa, United States of AmericaRobert W Burgess - The Jackson Laboratory, Bar Harbor, Maine, United States of America
- Resource Type
- Journal article
- Publication Details
- PLoS genetics, Vol.15(12), pp.e1008554-e1008554
- DOI
- 10.1371/journal.pgen.1008554
- PMID
- 31877124
- PMCID
- PMC6957209
- NLM abbreviation
- PLoS Genet
- ISSN
- 1553-7390
- eISSN
- 1553-7404
- Publisher
- United States
- Grant note
- R21 NS090030 / NINDS NIH HHS\r\nR01 NS055272 / NINDS NIH HHS
- Language
- English
- Date published
- 12/2019
- Academic Unit
- Neurology; Liberal Arts and Science Admin; Psychiatry; Iowa Neuroscience Institute; Biology
- Record Identifier
- 9983997988502771
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