In vivo encapsulation of nucleic acids using an engineered nonviral protein capsid |
| |
Authors: | Seth Lilavivat Debosmita Sardar Subrata Jana Geoffrey C Thomas Kenneth J Woycechowsky |
| |
Institution: | Department of Chemistry, University of Utah , 315 South 1400 East, Salt Lake City, Utah 84112, United States. |
| |
Abstract: | In Nature, protein capsids function as molecular containers for a wide variety of molecular cargoes. Such containers have great potential for applications in nanotechnology, which often require encapsulation of non-native guest molecules. Charge complementarity represents a potentially powerful strategy for engineering novel encapsulation systems. In an effort to explore the generality of this approach, we engineered a nonviral, 60-subunit capsid, lumazine synthase from Aquifex aeolicus (AaLS), to act as a container for nucleic acid. Four mutations were introduced per subunit to increase the positive charge at the inner surface of the capsid. Characterization of the mutant (AaLS-pos) revealed that the positive charges lead to the uptake of cellular RNA during production and assembly of the capsid in vivo. Surprisingly, AaLS-pos capsids were found to be enriched with RNA molecules approximately 200-350 bases in length, suggesting that this simple charge complementarity approach to RNA encapsulation leads to both high affinity and a degree of selectivity. The ability to control loading of RNA by tuning the charge at the inner surface of a protein capsid could illuminate aspects of genome recognition by viruses and pave the way for the development of improved RNA delivery systems. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|