PUBLICATION

Identification of maternal-effect genes in zebrafish using maternal crispants

Authors
Moravec, C.E., Voit, G.C., Otterlee, J., Pelegri, F.
ID
ZDB-PUB-210901-5
Date
2021
Source
Development (Cambridge, England)   148(19): (Journal)
Registered Authors
Pelegri, Francisco
Keywords
Crispr, Early development, Genome editing, Maternal-effect, Zebrafish, fhdc3, kpna7
MeSH Terms
  • Maternal Inheritance*
  • CRISPR-Cas Systems*
  • Ovum/cytology
  • Ovum/metabolism
  • Zebrafish Proteins/genetics*
  • Gene Editing/methods*
  • Animals
  • Zebrafish
(all 8)
PubMed
34463742 Full text @ Development
Abstract
In animals, early development is dependent on a pool of maternal factors, both RNA and proteins, which are required for basic cellular process and cell differentiation until zygotic genome activation. The role of a majority of these maternally expressed factors is not fully understood. By exploiting the biallelic editing ability of CRISPR-Cas9, we identify and characterize maternal-effect genes in a single generation, using a maternal crispant technique. We validated the ability to generate biallelic mutations in the germline by creating maternal crispants that phenocopied previously characterized maternal-effect genes: motley/birc5b, tmi/prc1l, and aura/mid1ip1. Additionally, by targeting maternally expressed genes of unknown function in zebrafish, we identified two new maternal-effect zebrafish genes, kpna7 and fhdc3. The genetic identity of these maternal crispants was confirmed by sequencing haploid progeny from F0 females, which allowed the analysis of newly induced lesions in the maternal germ line. Our studies show that maternal crispants allow for the effective identification and primary characterization of maternal-effect genes in a single generation, facilitating the reverse genetics analysis of maternal factors that drive embryonic development.
Genes / Markers
Figures
Figure Gallery (9 images)
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Expression
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
p4anua
    		Unknown
    uw107
      		Complex
      1 - 2 of 2
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      Human Disease / Model
      No data available
      Sequence Targeting Reagents
      Target Reagent Reagent Type
      birc5bCRISPR4-birc5bCRISPR
      birc5bCRISPR1-birc5bCRISPR
      birc5bCRISPR2-birc5bCRISPR
      birc5bCRISPR3-birc5bCRISPR
      fhdc3CRISPR2-fhdc3CRISPR
      fhdc3CRISPR3-fhdc3CRISPR
      fhdc3CRISPR4-fhdc3CRISPR
      fhdc3CRISPR1-fhdc3CRISPR
      kcns3bCRISPR1-kcns3bCRISPR
      kcns3bCRISPR2-kcns3bCRISPR
      1 - 10 of 28
      Show
      Fish
      No data available
      Antibodies
      Orthology
      No data available
      Engineered Foreign Genes
      No data available
      Mapping
      No data available
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      Citation
      Moravec, C.E., Voit, G.C., Otterlee, J., Pelegri, F. (2021) Identification of maternal-effect genes in zebrafish using maternal crispants. Development (Cambridge, England). 148(19).