Now, researchers have developed a revolutionary technique enabling live-cell protein tagging while maintaining protein function. This innovative approach, developed by a team led by Jenna N. Beyer and published in the journal Science, employs engineered molecular tools to enable precise editing of proteins within a short timeframe.
With this new technique, scientists can now tag proteins in real-time, allowing access to protein populations never before accessible on biologically relevant timescales. By using two splicing switches that they could program, scientists could start editing proteins only minutes after introducing donor proteins into the cells. This rapid editing capability marks a significant advancement over traditional tagging methods, which typically require longer periods and can compromise protein functionality.
Methodology and Mechanism
The protein editing system the research team developed functions like a splicing switch. This breakthrough allows for rapid prototyping of protein architectures. This approach incorporates NCrs into endogenous proteins. Consequently, it expands our ability to better understand protein dynamic function and interactome in the context of native live cellular biology.
The research project first validated this approach in a proof-of-principle study using model proteins, calnexin and β-actin. Calnexin is integral to the quality control of proteins within the endoplasmic reticulum, and β-actin is essential for cytoskeletal structure maintenance. Both proteins were successfully tagged, and their fluorescent signals persisted for several hours, allowing researchers to track their locations within the cell.
The efficiency of this technique is notable. Additionally, researchers found that splicing occurred within ten minutes following delivery of the donor protein. This speed is not often achieved by other tagging approaches. The fluorescent signals were exactly where we expected the target proteins to be, inside cells. This validation step helped to ensure our tagging process was correct.
Implications for Future Research
This powerful new approach paves the way for further, cutting-edge research in cell biology and biochemistry. This will allow researchers to make unparalleled observations of cellular processes in real-time without interference, leading to a deeper understanding of cellular processes. Including these noncanonical residues in their work will better enable scientists to explore new realms of protein function and interaction.
The possible uses of this technology are huge. It can be applied in many areas, such as pharmaceutical applications like drug development, disease modeling, and fundamental biological process understanding. These advances in tagging proteins in live cells ultimately empower researchers to conduct novel studies. This potent tool provides them with the means to disentangle complicated cellular processes.
“Intracellular protein editing enables incorporation of noncanonical residues in endogenous proteins.” – Jenna N. Beyer et al