Matthew Wilson
Last active: 3/28/2019

CRISPR/Cas9 engineering of a KIM-1 reporter human proximal tubule cell line.

Veach RA, Wilson MH
PLoS One. 2018 13 (9): e0204487

PMID: 30260998 · PMCID: PMC6160069 · DOI:10.1371/journal.pone.0204487

We used the CRISPR/Cas9 system to knock-in reporter transgenes at the kidney injury molecule-1 (KIM-1) locus and isolated human proximal tubule cell (HK-2) clones. PCR verified targeted knock-in of the luciferase and eGFP reporter at the KIM-1 locus. HK-2-KIM-1 reporter cells responded to various stimuli including hypoxia, cisplatin, and high glucose, indicative of upregulation of KIM-1 expression. We attempted using CRISPR/Cas9 to also engineer the KIM-1 reporter in telomerase-immortalized human RPTEC cells. However, these cells demonstrated an inability to undergo homologous recombination at the target locus. KIM-1-reporter human proximal tubular cells could be valuable tools in drug discovery for molecules inhibiting kidney injury. Additionally, our gene targeting strategy could be used in other cell lines to evaluate the biology of KIM-1 in vitro or in vivo.

MeSH Terms (16)

Acute Kidney Injury Cell Line Cisplatin CRISPR-Cas Systems Gene Knock-In Techniques Genes, Reporter Gene Targeting Genetic Engineering Glucose Green Fluorescent Proteins Hepatitis A Virus Cellular Receptor 1 Homologous Recombination Humans Kidney Tubules, Proximal Luciferases Up-Regulation

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