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CRISPR/Cas9 screening reveals key role of STK11/LKB1 in vasopressin-mediated regulation of Aqp2 transcription

GSE326336 Mus musculus (mouse) RNA-seq (gene expression) 44 samples Submitted 2026/03/30 Platform GPL34328
At a glance

RNA-seq (gene expression) · mouse · 44 samples

What it is
RNA-seq (gene expression)Gene activity measured by sequencing. The summary does not mention single-cell work, so this is most likely bulk: one average per sample, the usual starting point for comparing groups.
Organism
Mus musculus (mouse)
Samples
44 samples: 9 conditions, mostly with about 4 replicates eachGroups are read from the sample names, not from the paper. Check the list below.
Data you can get
A processed table (series matrix) and supplementary files from NCBI; the raw sequencing reads from ENA. Which of these hold counts depends on the study, so check the file list.
Summary (written by the submitters)
Identification of signaling networks is an essential goal in systems biology. Here, we use CRISPR knockout screening (employing a whole kinome sgRNA library) to identify functionally critical protein kinases in a well-studied Gs-dependent G-protein coupled receptor (GPCR)-signaling model, namely the vasopressin V2 receptor (V2R). Screening was done using a specially-designed fluorescence-based reporter cell line with green-fluorescent protein (GFP) co-transcribed with Aqp2, a gene whose transcription is dependent on vasopressin-mediated activation of protein kinase A (PKA).
Read the full summary ↓
The reporter line endogenously expresses V2R. Positive regulators (n=16) included PKA-catalytic subunit α (but not β). Mark2 (Par1b) and Dyrk1a (minibrain homolog). Negative regulators (n=13) included PKA-regulatory subunit type Iα, Stk11 (liver kinase B1 [LKB1]), and three TGF-β receptor subunits (Tgfbr1, Tgfbr2, Tgfbr3) (see https://esbl.nhlbi.nih.gov/Databases/Kinome-CRISPR-screen/ for full list). Dyrk1a and Stk11/LKB1 knockout cell lines were created for further study. Dyrk1a knockout cell lines failed to express AQP2 protein and exhibited a profound decrease in AQP2 mRNA. RNA-sequencing demonstrated widespread increases in cell-cycle transcripts, with a general defect in cell differentiation, accounting for AQP2 loss. Stk11/LKB1 knockout lines displayed marked increases in AQP2 protein and mRNA. RNA-sequencing and phospho-proteomic findings point to a signaling model in which Stk11/LKB1- and PKA-mediated phosphorylation events exert counteracting effects on the activities of cAMP-responsive transcriptional coactivator (CRTC) proteins. Finally, additional studies confirmed that TGF-β exposure to un-transformed cells results in a profound decrease in AQP2 mRNA abundance along with most recognized differentiation markers, consistent with our prior conclusion that TGF-β represses Aqp2 gene expression by inducing epithelial-mesenchymal transition.
This dataset
Open GSE326336 in finder → Find more mouse RNA-seq datasets →
Download · which file do I need?

Direct links to NCBI and ENA, no account and no request form. Pick by what you want to do: take the small processed table to look, the supplementary files to re-analyse, or the original raw data to start over.

1 · Just have a look

Processed table

The series matrix: one table of values per sample as the submitters processed it. The smallest download, and it opens in Excel or R. Start here to see what the data look like.

Series matrix ↓
2 · Re-analyse it

Supplementary files

The count tables and other files the authors uploaded. Take only the samples you need, or the whole study as GSE326336_RAW.tar (it can run to tens of gigabytes).

All supplementary files ↗ Whole study · RAW.tar ↓
3 · Start from the raw reads

Raw sequencing reads

The original FASTQ files, straight from the sequencer, held at ENA. Large, and you align them yourself, so only choose this if you need to re-process the study from scratch.

Raw reads on ENA ↗

What the files usually look like: For RNA-seq the counts are usually a table among the supplementary files (look for 'counts', 'expression', 'FPKM' or 'TPM' in the file name). The series matrix of a sequencing study is sometimes nearly empty.

Also filed as BioProject PRJNA1445127 and SRA study SRP687729. Searching any of these in the dataset finder brings you back here.

NCBI GEO page ↗
Study design
9 conditions, mostly with about 4 replicates each
WT_dDAVP_48h ×9 WT_veh_48h ×5 Stk11KO_veh_48h ×5 Stk11KO_dDAVP_48h ×5 Veh_Con ×4 Veh_dDAVP ×4 1Tgf_dDAVP ×4 10Tgf_dDAVP ×4 +1 more

Supports a between-group comparison across 44 samples.

How we read this: 9 replicated groups read from 44 sample titles; they account for 44 of them. These groups are guessed from the sample names, not taken from the paper, so check them against the sample list below before relying on them.

Samples in this study

+ 4 more — browse all 44 samples with per-sample file links →

Datasets on a similar topic

Search all mouse RNA-seq datasets in GEO →

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