0486598 - ÚMG 2018 RIV NL eng J - Článek v odborném periodiku
Kříž, Vítězslav - Krausová, Michaela - Burešová, Petra - Dobeš, Jan - Hrčkulák, Dušan - Babošová, Oľga - Švec, Jiří - Kořínek, Vladimír
Establishment of a tagged variant of Lgr4 receptor suitable for functional and expression studies in the mouse.
Transgenic Research. Roč. 26, č. 5 (2017), s. 689-701. ISSN 0962-8819. E-ISSN 1573-9368
Grant CEP: GA ČR(CZ) GA14-33952S; GA MŠMT LO1419; GA MŠMT(CZ) LM2015040; GA MŠMT ED2.1.00/19.0395
Klíčová slova: Genome editing * Hemagglutinin tag * Knock-in * R-spondin * TALENs * Wnt signaling
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 2.197, rok: 2017 ; AIS: 0.548, rok: 2017
DOI: https://doi.org/10.1007/s11248-017-0027-0

Leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) is produced in a broad spectrum of mouse embryonic and adult tissues and its deficiency results in embryonal or perinatal lethality. The LGR4 function was mainly related to its potentiation of canonical Wnt signaling, however, several recent studies associate LGR4 with additional signaling pathways. To obtain a suitable tool for studying the signaling properties of Lgr4, we generated a tagged variant of the Lgr4 receptor using gene targeting in the mouse oocyte. The modified Lgr4 allele expresses the Lgr4 protein fused with a triple hemagglutinin (3HA) tag located at the extracellular part of the protein. The allele is fully functional, enabling tracking of Lgr4 expression in the mouse tissues. We also show that via surface labeling, the 3HA tag allows direct isolation and analysis of living Lgr4-positive cells obtained from the small intestinal crypts. Finally, the HA tag-specific antibody can be employed to characterize the biochemical features of Lgr4 and to identify possible biding partners of the protein in cells derived from various mouse tissues.
Trvalý link: http://hdl.handle.net/11104/0281363