Protocol

Photoactivation-Based Labeling and In Vivo Tracking of RNA Molecules in the Nucleus

This protocol was adapted from "Photoactivation-Based Labeling and In Vivo Tracking of RNA Molecules in the Nucleus," Chapter 11, in Live Cell Imaging: A Laboratory Manual (eds. Goldman and Spector). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2005.

INTRODUCTION

This protocol describes a method for observing and measuring the movement of RNA molecules in the nucleus of living mammalian cells. Caged fluorescein-labeled DNA oligonucleotides are introduced into living mammalian cells, where they demonstrably hybridize to complementary RNA. After site-specific photoactivation at desired sites within the cell, the RNA movements away from those sites are followed and digitally recorded using a rapid acquisition microscopy system.