title: MARIS: method for analyzing RNA following intracellular sorting.
abstract: Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high quality RNA for transcriptome profiling following cellular fixation, intracellular immunofluorescent staining and FACS. MARIS can therefore be used to isolate high quality RNA from many otherwise inaccessible cell types simply based on immunofluorescent tagging of unique intracellular proteins. As proof of principle, we isolate RNA from sorted human embryonic stem cell-derived insulin-expressing cells as well as adult human β cells. MARIS is a basic molecular biology technique that could be used across several biological disciplines.
authors.affiliation: The Harvard Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, United States of America.
authors.forename: Siniša
authors.initials: S
authors.lastname: Hrvatin
authors.affiliation: The Harvard Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, United States of America.
authors.forename: Francis
authors.initials: F
authors.lastname: Deng
authors.affiliation: The Harvard Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, United States of America; Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Charles W
authors.initials: CW
authors.lastname: O'Donnell
authors.affiliation: Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: David K
authors.initials: DK
authors.lastname: Gifford
authors.affiliation: The Harvard Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, United States of America; Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, United States of America.
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Melton
mesh_terms: D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000737:chemistry; D005434:Flow Cytometry; D006801:Humans; D007515:Islets of Langerhans / Q000737:chemistry; D012313:RNA / Q000032:analysis*; D014158:Transcription, Genetic
chemical_list: D012313:RNA