Development of nanobody-based POLArIS orientation probes enabled multi-color/multi-target orientation imaging in living cells

In this study, we have developed new versions of POLArIS with nanobodies, which are compatible with animal immune libraries, and expanded color variations of POLArIS with cyan/green/yellow/red fluorescent proteins, enabling multi-color orientation imaging for multiple targets. Using nanobody-based POLArIS orientation probes, we performed two-color FPM of F-actin and vimentin in living cells. Furthermore, we made nanobody-based POLArIS probes that have different dipole orientations for adjusting the orientation of fluorescence polarization with respect to the target molecules. These nanobody-based POLArIS with options of colors and dipole orientations will enhance the performance of this probe for broader applications of fluorescence polarization imaging in living cells, tissues, and whole organisms.PMID:34090210 | DOI:10.1016/j.bbrc.2021.05.088

Source: Biochemical and Biophysical Research communications - June 5, 2021 Category: Biochemistry Authors: Nori Nakai Keisuke Sato Tomomi Tani Masahiko Kawagishi Hiromasa Ka Kenta Saito Sumio Terada Source Type: research