Livestock Gene Editing by One-step Embryo Manipulation.

Area of Science:

• Animal biotechnology
• Gene editing technologies
• Livestock improvement

Background:

• Clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) technology facilitates efficient gene editing in animals.
• Traditional CRISPR/Cas9 delivery to livestock embryos relies on microinjection.
• Generating gene-edited animals is crucial for agricultural advancements.

Purpose of the Study:

• To review the generation of gene-edited livestock via one-step embryo manipulation.
• To discuss various CRISPR/Cas9 delivery methods and reagent options.
• To highlight challenges and outcomes in livestock gene editing.

Main Methods:

• Review of current literature on CRISPR/Cas9 applications in livestock embryo editing.
• Comparison of intracytoplasmic microinjection and electroporation for CRISPR/Cas9 delivery.
• Analysis of different Cas9 and guide RNA (gRNA) reagent systems.

Main Results:

• Electroporation presents a viable alternative to microinjection for CRISPR/Cas9 delivery in livestock embryos.
• Mosaicism is a significant issue in one-step embryo editing, affecting gene-edited animal generation.
• Off-target mutations are infrequently observed in livestock generated through one-step editing.

Conclusions:

• One-step embryo manipulation is an effective strategy for generating gene-edited livestock.
• Optimizing delivery methods like electroporation and managing mosaicism are key for successful gene editing.
• CRISPR/Cas9 technology holds substantial promise for livestock breeding and research.