Live detection and purification of cells based on the expression of a histone chaperone, HIRA, using a binding peptide
| Autor: | Parvin Abraham, Aditi Majumder, Debasree Dutta, K. J. Kochurani, Madhumathy G. Nair, Annie A. Suganya, Jiss Maria Louis, K. Santhosh Kumar, Tessy Thomas Maliekal |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Cell Membrane Permeability
Aptamer Cellular differentiation Molecular Sequence Data Cell Cycle Proteins Cell Separation Stem cell marker Article Flow cytometry Mice medicine Animals Histone Chaperones Amino Acid Sequence Peptide sequence Cell potency Cells Cultured Multidisciplinary biology medicine.diagnostic_test Staining and Labeling Mouse Embryonic Stem Cells Flow Cytometry Peptide Fragments Histone P19 cell Biochemistry biology.protein Fluorescein-5-isothiocyanate Transcription Factors |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep17218 |
| Popis: | Flowcytometry 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. |
| Databáze: | OpenAIRE |
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