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Genetic compensation of γ CaMKII, an evolutionarily conserved gene

Sarah C Rothschild1, Sarah R Ingram1, Fu-I Lu2

  • 1Life Sciences, Virginia Commonwealth University, Richmond, VA 23284, USA.

Gene
|March 14, 2020
PubMed

Insights

CaMKII (calcium/calmodulin-dependent protein kinase) gene family is crucial for early zebrafish development. Genetic compensation mechanisms, like camk2g2 upregulation, occur when camk2g1 is disrupted.

Area of Science:

  • Developmental Biology
  • Genetics
  • Molecular Biology

Background:

  • Calcium/calmodulin-dependent protein kinase (CaMKII) genes are conserved across metazoans and vital from fertilization to adulthood.
  • The camk2g1 gene in zebrafish is implicated in early development, as shown by various experimental approaches.
  • Sequence conservation and gene duplication of CaMKII genes are observed in other teleost fish species.

Purpose of the Study:

  • To investigate the role of camk2g1 during early zebrafish development.
  • To explore the consequences of camk2g1 disruption using gene editing technologies.
  • To understand the transcriptional regulation and potential genetic compensation within the CaMKII gene family.

Main Methods:

  • Pharmacologic, dominant-negative, and antisense morpholino approaches in zebrafish.
  • Generation of a homozygous camk2g1 mutant using TALEN technology.
  • CRISPR/Cas9 gene editing in G0 zebrafish embryos to assess camk2g1 function.
  • Quantitative analysis of camk2 gene expression in mutant and wild-type embryos.

Main Results:

  • TALEN-generated camk2g1 mutants showed reproductive viability but a 4-fold upregulation of the paralog camk2g2, with other camk2 genes suppressed.
  • CRISPR/Cas9 G0 camk2g1 embryos exhibited phenotypes similar to morphants, including renal, aural, and ciliary defects.
  • These results suggest gene-specific transcriptional cross-talk and genetic compensation within the CaMKII gene family.

Conclusions:

  • The CaMKII gene family plays a significant role in early zebrafish development.
  • Genetic compensation, particularly the upregulation of camk2g2, is a key response to camk2g1 disruption.
  • CRISPR/Cas9 technology effectively recapitulates developmental phenotypes associated with camk2g1, validating its importance.
Keywords:
CRISPRCaMKIICiliaGenetic compensationKidneyTeleostZebrafish

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