"Thus, this new site-specific and cassette mutagenesis method is highly efficient, fast and versatile, likely resulting in its wide use for typical biomedical research, as well as for engineering and refining synthetic or mutant proteins from AI-assisted design."
BUFFALO, NY — November 6, 2025 — A new research paper was published in Volume 16 of Genes & Cancer on October 31, 2025, titled " P3a site-specific and cassette mutagenesis for seamless protein, RNA and plasmid engineering. "
In this study, Dr. Xiang-Jiao Yang from McGill University and McGill University Health Center developed an improved gene-editing technique that achieves nearly 100% success in creating precise DNA mutations. The method, called P3a mutagenesis, could significantly accelerate biomedical research and reduce costs by simplifying the engineering of proteins, RNAs, and plasmids used in studies of disease, genetics, and vaccine development.
Researchers have long relied on site-directed mutagenesis to study gene function, investigate disease-causing mutations, and develop therapeutic proteins. However, existing techniques often struggle with inefficiency, long processing times, and high error rates, especially when editing large DNA molecules. The newly developed P3a method addresses these challenges by using specially designed primers combined with high-fidelity enzymes, enabling fast and accurate DNA editing.
"In summary, we have developed a fast, economical and highly efficient site-specific mutagenesis method based on Q5 and SuperFi II high-fidelity DNA polymerases, along with primer pairs with 3′-overhangs."
The study shows that P3a mutagenesis can efficiently introduce a wide range of genetic modifications, including single-point mutations, large deletions, and insertions, into DNA fragments up to 13.4 kilobases long. It also allows for precise editing of key biomedical targets, such as cancer-related genes, variants linked to neurodevelopmental disorders, and spike proteins from evolving SARS-CoV-2 strains. Unlike older approaches that often require multiple steps or complex cloning procedures, the P3a method delivers correct edits within a few days and with minimal laboratory resources.
Thanks to its reliability and versatility, P3a mutagenesis is positioned to become a powerful tool in modern bioscience. It simplifies the analysis of disease-related mutations, supports the rapid testing of synthetic and AI-designed proteins, and enables seamless modification of genome-editing systems like CRISPR-Cas9. The method also provides a cost-effective way to engineer viral variants, such as those linked to COVID-19, offering a practical tool for ongoing pandemic response efforts.
By significantly improving the efficiency of genetic engineering while reducing both time and cost, this new method is expected to be widely adopted in biomedical research. It offers scientists a flexible, precise, and scalable solution for studying genes and proteins in greater depth and with unprecedented speed.
Continue reading: https://doi.org/10.18632/genesandcancer.243
