Xenopus laevis transgenesis by sperm nuclear injection

Area of Science:

• Developmental Biology
• Genetics
• Molecular Biology

Background:

• Stable integration of transgenes is crucial for creating genetically modified organisms.
• Xenopus laevis is a valuable model organism for developmental studies.

Purpose of the Study:

• To describe a novel, efficient procedure for stable transgene integration in Xenopus laevis embryos.
• To establish a method for generating transgenic reporter frog lines.

Main Methods:

• Linear DNA containing the transgene was incorporated into detergent-treated sperm nuclei.
• Microinjection of modified sperm nuclei into unfertilized eggs.
• Screening of resulting viable embryos for transgene activity.

Main Results:

• Successful stable integration of transgenes into Xenopus laevis embryos.
• Transgene integration rates ranged from 10% to 40% in surviving embryos.
• Routine germline transmission of integrated transgenes was achieved.

Conclusions:

• The described method provides a robust approach for generating transgenic Xenopus.
• This technique is well-suited for creating transgenic reporter lines for various research applications.