Navigating gene editing in porcine embryos: Methods, challenges, and future perspectives

Area of Science:

• Biotechnology
• Genetics
• Animal Science

Background:

• Gene editing technologies, particularly CRISPR/Cas9, are revolutionizing genetic modification.
• Porcine embryos are increasingly utilized in biomedical and agricultural research due to these advancements.

Purpose of the Study:

• To comprehensively review gene editing methodologies and delivery methods in porcine embryos.
• To evaluate the advantages and limitations of different biological sources (in vivo vs. in vitro oocytes/embryos).
• To discuss innovative approaches like sperm-mediated gene transfer (SMGT) and testis-mediated gene transfer (TMGT).

Main Methods:

• Review of gene editing techniques including CRISPR/Cas9.
• Examination of delivery methods: somatic cell nuclear transfer (SCNT), microinjection, electroporation, and lipofection.
• Analysis of germline manipulation strategies (SMGT, TMGT).

Main Results:

• CRISPR/Cas9 significantly advances genetic modification in porcine embryos.
• Various delivery methods and biological sources have distinct advantages and limitations.
• Innovative germ cell manipulation techniques offer new avenues for genetic modification.

Conclusions:

• Current gene editing strategies face challenges such as off-target events and mosaicism.
• Refinement of methods is crucial to reduce mosaicism and enhance editing accuracy.
• Further advancements are necessary to fully realize the potential of gene editing in agriculture and biomedicine.