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Related Concept Videos

CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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AP-4 loss in CRISPR-edited zebrafish affects early embryo development.

Olivia G Pembridge1, Natalie S Wallace2, Thomas P Clements1

  • 1Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA.

Advances in Biological Regulation
|January 15, 2023
PubMed
Summary

Mutations in adaptor protein 4 (AP-4) cause neurological disorders. Zebrafish gene editing reveals AP-4 loss impacts embryo development and autophagy gene expression, offering a new model for studying these conditions.

Keywords:
CRISPRCoat proteinsGene editingMembrane traffickingZebrafish

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Area of Science:

  • Cell Biology
  • Developmental Biology
  • Genetics

Background:

  • Mutations in adaptor protein 4 (AP-4) are linked to severe neurological disorders, including spastic paraplegia, microcephaly, and intellectual disabilities.
  • AP-4's role in membrane trafficking, particularly its cargo ATG9A (an autophagy protein), is crucial, but organismal-level understanding is limited due to gene loss in common model systems.
  • Zebrafish (Danio rerio) possess conserved AP-4 genes, making them a suitable model for studying AP-4's developmental and neurological functions.

Purpose of the Study:

  • To investigate the effects of AP-4 and tepsin gene loss on zebrafish embryo development using CRISPR-ExoCas9 gene editing.
  • To explore the impact of AP-4 single gene knockouts on the expression of autophagy-related genes (atg9a and map1lc3b) in zebrafish embryos.
  • To establish zebrafish as a model organism for dissecting AP-4's function in membrane trafficking and autophagy.

Main Methods:

  • CRISPR-ExoCas9 gene editing was employed to create single gene knockouts for AP-4 and tepsin in zebrafish embryos.
  • Zebrafish embryos were analyzed at 24 hours post-fertilization (hpf) for developmental abnormalities.
  • Quantitative analysis of atg9a and map1lc3b gene expression was performed in gene-edited zebrafish embryos.

Main Results:

  • Single gene editing of AP-4 or tepsin resulted in observable abnormal head morphology and neural necrosis in zebrafish embryos at 24 hpf.
  • The study provides the first evidence of altered expression patterns for autophagy genes atg9a and map1lc3b following AP-4 gene knockout in zebrafish embryos.
  • These findings highlight the critical role of AP-4 in early embryonic development and neurogenesis.

Conclusions:

  • Zebrafish gene editing effectively models the developmental consequences of AP-4 dysfunction, mirroring aspects of human neurological disorders.
  • AP-4 plays a significant role in regulating autophagy gene expression during early development.
  • Zebrafish represent a valuable and adaptable model system for future research into AP-4-mediated membrane trafficking and autophagy in the context of complex diseases.