COLUMN FRACTION SCREENING OF CPMV PARTICLES

这个子项目是众多研究子项目之一

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There is a need to be able to separate and characterize the RNA-filled, empty, and aggregated fractions following UV treatment in order to purify the RNA-containing, non-infectious single CPMV particles for further in vivo studies. In order to show whether the many column fractions have these physical characteristics, the goal is to be able to visualize fractions collected from columns or sucrose gradients in an efficient manner. Since a typical column run yields between 20-100 fractions, the preparation of individual CPMV fractions for EM would benefit significantly from automation. Analysis of the TEM images would then be needed, to view the particles in each fraction to identify and quantify the proportion of RNA-filled capsids, empty capsids, aggregated debris, and broken particles. This procedure would make the validation of individual inactivated lots of CPMV nanoparticles feasible for in vivo use.