The IMPDH1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from human colorectal adenocarcinoma HT29 cells, with targeted disruption of the IMPDH1 gene. Generated via CRISPR/Cas9-mediated gene disruption, this heterogeneous knockout pool abrogates IMPDH1 protein function, providing a powerful tool to study de novo guanine nucleotide biosynthesis and its roles in proliferation and signaling.
The HT29 parental cell line originated from a poorly differentiated human colorectal adenocarcinoma and is a well-established model for colorectal cancer. These adherent epithelial cells can undergo enterocyte-like differentiation in glucose-free medium, with upregulated mucin secretion and tight junction-mediated barrier formation. This inducible differentiation, combined with a tumorigenic background, makes HT29 cells ideal for studying the interplay between proliferation, differentiation, and epithelial barrier integrity.
IMPDH1 encodes inosine monophosphate dehydrogenase type I, catalyzing the rate-limiting NAD-dependent conversion of IMP to XMP in de novo guanine nucleotide synthesis. The enzyme is allosterically inhibited by GTP/GDP and transcriptionally controlled by MYC, PI3K/AKT/mTOR, and p53. IMPDH1 activity determines intracellular GTP/GDP pools, which regulate small GTPases (Ras, Rho, Arf) and sustain DNA/RNA synthesis and cell cycle progression. IMPDH1 dynamically assembles into filaments with IMPDH2 and interacts with CTPS and polysomal mRNA, linking nucleotide metabolism to cytoophidia formation and translational control.
IMPDH1 knockout in HT29 cells depletes guanine nucleotide pools, impairing proliferation and GTP-dependent signaling while potentially altering differentiation. The HT29 line??s capacity for enterocyte maturation and barrier formation makes this model uniquely suited to examine how nucleotide metabolism influences mucin production and epithelial polarity. Moreover, knockout cells exhibit heightened sensitivity to antimetabolites like mycophenolic acid and ribavirin, facilitating drug response studies and resistance profiling.
This polyclonal knockout pool supports diverse applications: proliferation assays (MTT, BrdU), nucleotide quantification (HPLC/LC-MS), GTP loading assays, and cell cycle analysis by flow cytometry. IMPDH1 loss can be validated by Western blot, RT-qPCR, and immunofluorescence for cytoophidia, with downstream functional studies using RNA-seq, migration/invasion assays, and differentiation protocols to assess barrier function and mucin secretion. For further information or technical assistance, please contact Ascent Research.
