A study reports that a 2A peptide derived from porcine teschovirus-1 (P2A) exhibits the highest cleavage efficiency among four 2A peptides (P2A, T2A, E2A, and F2A) in human cell lines, zebrafish embryos, and mouse liver. The researchers constructed expression plasmids containing these 2A peptides and evaluated their cleavage efficiency using Western blotting and confocal microscopy. The results showed that P2A had the highest cleavage efficiency in all tested contexts. The study also developed cloning vectors with a simian cytomegalovirus (sCMV) promoter and a 2A gene flanked by multiple cloning sites, which can be used to simultaneously express multiple genes in mammalian cells, zebrafish, and mice. The findings suggest that P2A is a promising candidate for use in biomedical research due to its high cleavage efficiency and the availability of cloning vectors that facilitate its application. The study highlights the importance of selecting the appropriate 2A peptide for efficient gene expression in different biological systems.A study reports that a 2A peptide derived from porcine teschovirus-1 (P2A) exhibits the highest cleavage efficiency among four 2A peptides (P2A, T2A, E2A, and F2A) in human cell lines, zebrafish embryos, and mouse liver. The researchers constructed expression plasmids containing these 2A peptides and evaluated their cleavage efficiency using Western blotting and confocal microscopy. The results showed that P2A had the highest cleavage efficiency in all tested contexts. The study also developed cloning vectors with a simian cytomegalovirus (sCMV) promoter and a 2A gene flanked by multiple cloning sites, which can be used to simultaneously express multiple genes in mammalian cells, zebrafish, and mice. The findings suggest that P2A is a promising candidate for use in biomedical research due to its high cleavage efficiency and the availability of cloning vectors that facilitate its application. The study highlights the importance of selecting the appropriate 2A peptide for efficient gene expression in different biological systems.