Laser-assisted Cytoplasmic Microinjection in Livestock Zygotes

Overview

This protocol demonstrates the technique of cytoplasmic microinjection in farm animal zygotes, enabling the delivery of various solutions, including genome editing tools, into one-cell embryos.

Key Study Components

Area of Science

• Animal Biotechnology
• Transgenesis
• Genome Editing

Background

• Cytoplasmic microinjection is a method used to introduce substances into zygotes.
• This technique is particularly useful in livestock research.
• Genome editing tools like CRISPR Cas9 can be delivered to create knockout animals.
• Efficient injection minimizes adverse effects on animal health.

Purpose of Study

• To effectively deliver solutions into the cytoplasm of livestock zygotes.
• To facilitate the creation of transgenic animals for research purposes.
• To improve the reliability of genome editing in large animals.

Methods Used

• Preparation of injection micropipette using a borosilicate glass capillary.
• Utilization of a micropipette puller to achieve a thin tip with a long taper.
• Application of a blunt-end needle for microinjection.
• Use of laser assistance to enhance injection precision.

Main Results

• The method allows for the successful delivery of solutions into zygotes.
• High injection efficiency was observed with minimal adverse effects.
• Demonstrated potential for creating genetically modified livestock.
• Provides a reliable approach for transgenesis in large animals.

Conclusions

• Cytoplasmic microinjection is an effective technique for genome editing in livestock.
• This method enhances the study of gene function in large animals.
• It offers a straightforward approach to producing transgenic animals.

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