title: Live detection and purification of cells based on the expression of a histone chaperone, HIRA, using a binding peptide.
abstract: Flow cytometry is a reliable method for identification and purification of live cells from a heterogeneous population. Since permeabilized cells cannot be sorted live in a FACS sorter, its application in isolation of functional cells largely depends on antibodies for surface markers. In various fields of biology we find intracellular markers that reveal subpopulations of biological significance. Cell cycle stage specific molecules, metastatic signature molecules, stemness associated proteins etc. are examples of potential markers that could improve the research and therapy enormously. Currently their use is restricted by lack of techniques that allow live detection. Even though a few methods like aptamers, droplet-based microfluidics and smartflares are reported, their application is limited. Here, for the first time we report a simple, cost-effective and efficient method of live sorting of cells based on the expression of an intracellular marker using a fluorophore-tagged binding peptide. The target molecule selected was a histone chaperone, HIRA, the expression of which can predict the fate of differentiating myoblast. Our results confirm that the peptide shows specific interaction with its target; and it can be used to separate cells with differential expression of HIRA. Further, this method offers high purity and viability for the isolated cells.
authors.affiliation: Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India, 695014.
authors.forename: K J
authors.initials: KJ
authors.lastname: Kochurani
authors.affiliation: Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India, 695014.
authors.forename: Annie A
authors.initials: AA
authors.lastname: Suganya
authors.affiliation: Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India, 695014.
authors.forename: Madhumathy G
authors.initials: MG
authors.lastname: Nair
authors.affiliation: Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India, 695014.
authors.forename: Jiss Maria
authors.initials: JM
authors.lastname: Louis
authors.affiliation: Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India, 695014.
authors.forename: Aditi
authors.initials: A
authors.lastname: Majumder
authors.affiliation: Chemical Biology Group, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India, 695014.
authors.forename: Santhosh K
authors.initials: SK
authors.lastname: Kumar
authors.affiliation: Chemical Biology Group, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India, 695014.
authors.forename: Parvin
authors.initials: P
authors.lastname: Abraham
authors.affiliation: Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India, 695014.
authors.forename: Debasree
authors.initials: D
authors.lastname: Dutta
authors.affiliation: Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India, 695014.
authors.forename: Tessy T
authors.initials: TT
authors.lastname: Maliekal
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002463:Cell Membrane Permeability; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D056488:Histone Chaperones / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D000066450:Mouse Embryonic Stem Cells / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D013194:Staining and Labeling; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; C484187:Hira protein, mouse; D056488:Histone Chaperones; D010446:Peptide Fragments; D014157:Transcription Factors; D016650:Fluorescein-5-isothiocyanate