Newswise — Chicago (April 14, 2025) — Researchers have developed a new CRISPR-based method that makes it quicker and easier to create hybrid proteins. The new approach could be useful for creating building blocks for a variety of synthetic systems, making genetically encoded biosensors and creating proteins that carry out multiple functions.
CRISPR is used to edit DNA, allowing scientists to fix genetic mistakes, study diseases and even develop new treatments. However, finding workable DNA insertion sites in proteins has traditionally been a slow and expensive process, making it challenging to use CRISPR for high-throughput applications.
With the new CRISPR-based method, called PAM scanning, researchers can quickly identify safe sites where new code can be added without disrupting the protein’s normal folding or function.
The study’s first author Jacinda Pujols, a doctoral candidate at the University of Miami Miller School of Medicine, will present the research at the , which is being held April 12–15 in Chicago.
“Using PAM scanning, we created hundreds of functional chimeras — hybrid proteins created by fusing domains from different proteins,” said Pujols. “The high-throughput potential of this CRISPR-based system could enable rapid testing of emerging pathogen variants and inhibitor resistance, paving the way for faster antiviral therapeutic development.”
Pujols conceived the PAM scanning method when looking for a way to address the urgent need for rapid antiviral screening during the COVID-19 pandemic. She had the idea to design hybrid proteins by using a custom GPCR signaling pathway that contained special sites to which viral proteases could bind. If a protease was present, it would cut the site, shutting down the signal. A protease inhibitor could then be used to turn the signal back on. If the signaling turned off and on as expected, it would indicate that inserting a new sequence at that site wouldn’t interfere with protein folding or function.
Pujols implemented her idea in a model yeast system, which is commonly used in synthetic biology. She also developed a way to combine pooled experiments with cell sorting to rapidly build and test large numbers of chimeras.
“This method is broadly applicable, including for modifying human proteins,” said Pujols. “For example, it could also be used to quickly construct synthetic receptors for therapies similar to CAR T-cell treatments, which modify a patient's own T-cells to better recognize and attack cancer cells.”
The researchers say that integrating PAM scanning with AI-guided protein design could further enhance functional chimera creation, making it possible to develop a wide range of proteins tailored to specific needs by combining different functional components.
Jacinda Pujols will present this research from 4:30 to 6:30 p.m. CDT on Monday, April 14, at the Lakeside Center of the McCormick Place Convention Center. Contact the media team for more information.
About the American Society for Biochemistry and Molecular Biology (ASBMB)
The ASBMB is a nonprofit scientific and educational organization with more than 12,000 members worldwide. Founded in 1906 to advance the science of biochemistry and molecular biology, the society publishes three peer-reviewed journals, advocates for funding of basic research and education, supports science education at all levels, and promotes the diversity of individuals entering the scientific workforce.
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