An Endogenously Tagged Fluorescent Fusion Protein Library in Mouse Embryonic Stem Cells

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F17%3A00485511" target="_blank" >RIV/68081707:_____/17:00485511 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.stemcr.2017.08.022" target="_blank" >http://dx.doi.org/10.1016/j.stemcr.2017.08.022</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.stemcr.2017.08.022" target="_blank" >10.1016/j.stemcr.2017.08.022</a>

Jazyk výsledku

angličtina

Název v původním jazyce

An Endogenously Tagged Fluorescent Fusion Protein Library in Mouse Embryonic Stem Cells

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

An Endogenously Tagged Fluorescent Fusion Protein Library in Mouse Embryonic Stem Cells

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Stem Cell Reports

ISSN

2213-6711

e-ISSN

—

Svazek periodika

9

Číslo periodika v rámci svazku

4

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

11

Strana od-do

1304-1314

Kód UT WoS článku

000412660000024

EID výsledku v databázi Scopus

—