CRISPR/Cas9-mediated tryptophan hydroxylase 1 knockout decreases calcium transportation in goat mammary epithelial cells
نویسندگان
چکیده
Calcium is one of the major mineral nutrients in goat milk. Tryptophan hydroxylase 1 (TPH1) a rate-limiting enzyme catalyzing hydroxylation l-tryptophan into 5-hydroxytryptamine (5-HT, serotonin) essential for maintaining calcium homeostasis. In this study, TPH1 knockout mammary epithelial cells(GMEC) heterozygous clone with no off-target effects were obtained after transfection Cas9/sgRNA expression vector firstly. Then content 5-HT, intracellular level, and abundance genes related to transportation was evaluated compared wild-type GMEC, explore impact on transportation, respectively. Wild-type GMEC further treated exogenous 5-HTP confirm role regulating homeostasis GMEC. The 5-HT synthesis level decreased gene mRNA secretory-pathway Ca2+-ATPase1 (SPCA1) plasma membrane (PMCA1) up-regulated while Ca2+-ATPase2 (SPCA2) down-regulated Up-regulation parathyroid hormone-related peptide (PTHrP), key regulator metabolism, induced by blocked knockout. Results suggested that via PTHrP transportation-related factors
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ژورنال
عنوان ژورنال: Biochemical Engineering Journal
سال: 2021
ISSN: ['1873-295X', '1369-703X']
DOI: https://doi.org/10.1016/j.bej.2021.108015