Nuclear transcriptomes at high resolution using retooled INTACT
Reynoso et al. describe the Isolation of Nuclei from TAgged specific Cell Types (INTACT) in rice. In this method, modified from one described previously in Arabidopsis and tomato, nuclei are labeled with a biotinylated nuclear-envelope anchored protein. Best results were obtained when a rice WIP (WPP-Interacting protein) was fused to GFP and BLRP (biotin ligase target peptide). The protein is biotinylated at the outer surface of the nucleus through a second transgene encoding biotin ligase. Biotin-labeled nuclei are then purified through the use of streptavidin-coated beads. The purified nuclei are sources of nuclear RNA and proteins for studies of gene regulation. Plant Physiol.
