Autor: |
Arigela Harikumar, Raghu Ram Edupuganti, Matan Sorek, Gajendra Kumar Azad, Styliani Markoulaki, Petra Sehnalová, Soňa Legartová, Eva Bártová, Shlomit Farkash-Amar, Rudolf Jaenisch, Uri Alon, Eran Meshorer |
Jazyk: |
angličtina |
Rok vydání: |
2017 |
Předmět: |
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Zdroj: |
Stem Cell Reports, Vol 9, Iss 4, Pp 1304-1314 (2017) |
Druh dokumentu: |
article |
ISSN: |
2213-6711 |
DOI: |
10.1016/j.stemcr.2017.08.022 |
Popis: |
Embryonic stem cells (ESCs), with their dual capacity to self-renew and differentiate, are commonly used to study differentiation, epigenetic regulation, lineage choices, and more. Using non-directed retroviral integration of a YFP/Cherry exon into mouse ESCs, we generated a library of over 200 endogenously tagged fluorescent fusion proteins and present several proof-of-concept applications of this library. We show the utility of this library to track proteins in living cells; screen for pluripotency-related factors; identify heterogeneously expressing proteins; measure the dynamics of endogenously labeled proteins; track proteins recruited to sites of DNA damage; pull down tagged fluorescent fusion proteins using anti-Cherry antibodies; and test for interaction partners. Thus, this library can be used in a variety of different directions, either exploiting the fluorescent tag for imaging-based techniques or utilizing the fluorescent fusion protein for biochemical pull-down assays, including immunoprecipitation, co-immunoprecipitation, chromatin immunoprecipitation, and more. |
Databáze: |
Directory of Open Access Journals |
Externí odkaz: |
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