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Characterization of in vitro models of SLC30A10 deficiency

Biometals. 2021-03; 
Milankumar Prajapati, Michael A Pettiglio, Heather L Conboy, Courtney J Mercadante, Shintaro Hojyo, Toshiyuki Fukada, Thomas B Bartnikas
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Gene Synthesis … All experiments were carried out using cultures in the log phase of growth. Generation of SLC30A10 mutant Hep3B cells. CRISPR/Cas9-mediated biallelic SLC30A10 mutation was performed by GenScript (Piscataway, NJ) … Get A Quote

摘要

Manganese (Mn), an essential metal, can be toxic at elevated levels. In 2012, the first inherited cause of Mn excess was reported in patients with mutations in SLC30A10, a Mn efflux transporter. To explore the function of SLC30A10 in vitro, the current study used CRISPR/Cas9 gene editing to develop a stable SLC30A10 mutant Hep3B hepatoma cell line and collagenase perfusion in live mice to isolate primary hepatocytes deficient in Slc30a10. We also compared phenotypes of primary vs. non-primary cell lines to determine if they both serve as reliable in vitro models for the known physiological roles of SLC30A10. Mutant SLC30A10 Hep3B cells had increased Mn levels and decreased viability when exposed to excess Mn. T... More

关键词

CRISPR-Cas9, Hep3B, Liver, Manganese, Primary hepatocytes, SLC30A10, SLC39A14, Toxicity, Whole genome sequencing
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