| First Author | Harikumar A | Year | 2017 |
| Journal | Stem Cell Reports | Volume | 9 |
| Issue | 4 | Pages | 1304-1314 |
| PubMed ID | 28966122 | Mgi Jnum | J:255088 |
| Mgi Id | MGI:6108958 | Doi | 10.1016/j.stemcr.2017.08.022 |
| Citation | Harikumar A, et al. (2017) An Endogenously Tagged Fluorescent Fusion Protein Library in Mouse Embryonic Stem Cells. Stem Cell Reports 9(4):1304-1314 |
| abstractText | 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. |
