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VitelloTag: a tool for high-throughput cargo delivery into oocytes.

D Nathaniel Clarke1, Akshay Kane2, Margherita Perillo2

  • 1Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142,USA.

Development (Cambridge, England)
|August 22, 2024
PubMed
Summary

VitelloTag offers a cost-effective, high-throughput method for delivering molecular tools into oocytes, overcoming limitations of traditional microinjection. This novel system utilizes vitellogenin-derived fusion proteins for efficient gene editing in developmental and reproductive biology research.

Keywords:
CRISPR/Cas9Gene editingOocyte deliveryTransgenesisVitellogenesis

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Area of Science:

  • Developmental Biology
  • Reproductive Biology
  • Molecular Biology
  • Biotechnology

Background:

  • Delivering molecular tools into oocytes is crucial for research in developmental and reproductive biology.
  • Conventional microinjection is labor-intensive, requires specialized equipment, and has low efficiency, especially for delicate oocytes or species with limited spawning seasons.

Purpose of the Study:

  • To develop a novel, cost-effective, and high-throughput system for delivering molecular tools into oocytes.
  • To overcome the limitations associated with traditional microinjection techniques in oocyte research.

Main Methods:

  • Development of VitelloTag, a system utilizing vitellogenin-derived fusion proteins.
  • Utilizing receptor-mediated endocytosis for efficient cargo delivery into oocytes.
  • Application of VitelloTag for delivering Cas9/sgRNA complexes for gene knockout.

Main Results:

  • VitelloTag provides a cost-effective and high-throughput solution for oocyte delivery.
  • The system enables efficient cargo delivery via receptor-mediated endocytosis.
  • Successful gene knockout was demonstrated in two distantly related species using VitelloTag.

Conclusions:

  • VitelloTag represents a significant advancement in oocyte delivery methods for developmental and reproductive biology.
  • The system offers a practical and efficient alternative to microinjection, broadening research possibilities.
  • VitelloTag facilitates high-throughput gene editing in oocytes across diverse species.