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Related Experiment Videos

'Artificial spermatid'-mediated genome editing†.

Lingbo Wang1,2, Jinsong Li1

  • 1State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, China.

Biology of Reproduction
|May 12, 2019
PubMed
Summary
This summary is machine-generated.

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Researchers developed artificial spermatids using genetically modified haploid embryonic stem cells (haESCs). This breakthrough enables efficient generation of genetically modified mice, overcoming limitations of traditional sperm-mediated gene transfer methods.

Area of Science:

  • Reproductive Biology
  • Developmental Biology
  • Genetics

Background:

  • Sperm-mediated gene transfer (SMGT) has limitations in producing stable and diverse genetic modifications in offspring.
  • Haploid embryonic stem cells (haESCs) can be derived from haploid embryos and maintain pluripotency and haploidy.
  • Androgenetic haploid ESCs (AG-haESCs) can generate semi-cloned (SC) mice, carrying only the paternal genome.

Purpose of the Study:

  • To develop a more efficient and versatile method for generating genetically modified animals.
  • To overcome the limitations of sperm-mediated gene transfer (SMGT).
  • To establish 'artificial spermatids' for creating genetically modified offspring with desired traits.

Main Methods:

  • Derivation of haploid embryonic stem cells (haESCs) from androgenetic haploid blastocysts (AG-haESCs).
Keywords:
CRISPR-Cas9androgenetic haESCsgene editinghaploid embryonic stem cellssemi-cloned mouse (SC mouse)‘artificial spermatids’

Related Experiment Videos

  • Genetic modification of AG-haESCs, specifically the double knockout (DKO) of H19-DMR and IG-DMR.
  • Generation of semi-cloned (SC) mice by injecting modified DKO-AG-haESCs into oocytes.
  • Utilizing CRISPR-Cas technology for in vitro gene modifications of DKO-AG-haESCs.

    • Main Results:

    • Removal of H19-DMR and IG-DMR in AG-haESCs (DKO-AG-haESCs) enabled stable and efficient production of SC animals.
    • DKO-AG-haESCs served as effective 'artificial spermatids', supporting multiple rounds of in vitro genetic modification.
    • Efficient generation of live, fertile SC mice with desired genetic modifications was achieved.
    • This method provides a versatile genetic tool for in vivo gene function analysis.

    Conclusions:

    • DKO-AG-haESCs, termed 'artificial spermatids', offer a superior alternative to traditional SMGT for creating genetically modified animals.
    • This approach allows for efficient, in vitro genetic manipulation before generating offspring.
    • The technology facilitates the creation of genetically modified mice for diverse research applications, including in vivo gene function studies.