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To investigate the variability of aquaporin-2 expression in collecting duct cells and its regulatory mechanisms.

GFP Reporter System Reveals Cell-to-Cell Variability in Aquaporin-2 Expression

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To investigate the variability of aquaporin-2 expression in collecting duct cells and its regulatory mechanisms.
Engineered Aqp2 reporter cell line using CRISPR/Cas9 to insert GFP at Aqp2 locus.
Tested effects of dDAVP and forskolin on GFP and Aqp2 expression.
Flow-sorted GFP-positive and GFP-negative cells for further analysis.
Conducted RNA-seq analysis on GFP-positive and GFP-negative populations.
dDAVP stimulation increased GFP and Aqp2 expression, with variability in expression across cells.
Less than 50% of cells showed detectable GFP and Aqp2 expression post-stimulation.
Flow cytometry revealed differences in cell cycle distribution between GFP+ and GFP- cells.
RNA-seq showed higher abundance of cell-cycle related transcripts in GFP- cells.

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Demonstrates variability in aquaporin-2 expression in engineered cell lines, suggesting important transcriptional regulation mechanisms.

Chen et al. (2026) studied this question.

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