A transgenic-cloned pig model expressing non-fluorescent modified Plum

Area of Science:

• Biotechnology
• Immunology
• Genetics

Background:

• Genetically modified pigs expressing fluorescent proteins are vital biomedical research tools.
• Xeno-antigenicity of fluorescent proteins can impede engraftment and survival in transplantation studies.
• A need exists for pigs immunologically tolerant to fluorescent protein antigens.

Purpose of the Study:

• To generate a transgenic (Tg)-cloned pig that is immunologically tolerant to fluorescent protein antigens.
• To overcome the limitations of xeno-antigenicity in fluorescent protein-based research.

Main Methods:

• Generated a Tg-cloned pig expressing a modified Plum (mPlum) fluorescent protein derivative.
• Utilized a single amino acid substitution in the mPlum chromophore.
• Confirmed antigenicity using western blot and immunohistochemistry.

Main Results:

• The Tg-cloned pig's cells and tissues did not exhibit fluorescence.
• Western blot and immunohistochemistry confirmed mPlum retained the same antigenicity as Plum.
• Demonstrated proof of principle for creating immunologically tolerant cloned pigs.

Conclusions:

• Successfully generated a Tg-cloned pig expressing a non-fluorescent but antigenic mPlum protein.
• Established a foundation for creating cloned pigs immunologically tolerant to fluorescent protein antigens.
• This work facilitates advancements in transplantation research using fluorescent markers.