The GPSM2 Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the GPSM2 gene has been disrupted. This heterogeneous population retains the polyclonal nature of the editing outcome, providing a robust loss-of-function model for studying GPSM2-dependent processes without selection for a single clonal genotype.
The HT29 cell line is derived from a human colorectal adenocarcinoma and exhibits epithelial morphology with the capacity for spontaneous goblet cell differentiation under appropriate culture conditions. Widely employed as an intestinal epithelial model, HT29 cells are instrumental in investigations of colorectal cancer biology, epithelial barrier function, and drug transport mechanisms.
GPSM2 functions as a scaffold protein that integrates G protein and polarity signals at the cell cortex to regulate mitotic spindle orientation and asymmetric cell division. It directly binds inactive G??i/o subunits, recruiting the NuMA-dynein complex to generate pulling forces on astral microtubules, and its activity is modulated by mitotic kinases such as Aurora A and polarity proteins including aPKC. Downstream, GPSM2 orchestrates interactions with Inscuteable, Par3, and adherens junction components such as PLEKHA7, bridging spindle positioning with planar cell polarity and Wnt/??-catenin pathway outputs.
In the colorectal adenocarcinoma setting, GPSM2-mediated control of spindle orientation and cell polarity is pivotal for maintaining epithelial architecture and regulating symmetric versus asymmetric divisions, processes frequently dysregulated in tumor initiation and progression. Disruption of GPSM2 in HT29 cells therefore offers a valuable tool to dissect the contribution of altered G protein signaling, spindle misorientation, and polarity defects to colorectal cancer cell behavior, drug responsiveness, and stem cell dynamics.
Typical experimental applications of these polyclonal knockout cells include immunofluorescence-based analysis of spindle orientation defects, colony formation and migration assays to evaluate tumorigenic potential, cell cycle flow cytometry, and RNA-seq transcriptomic profiling to uncover molecular signatures downstream of GPSM2 loss. Additionally, they serve as a robust platform for drug sensitivity screenings and differentiation assays pertinent to intestinal epithelial biology and colorectal cancer therapy. For further information or custom cell engineering services, please contact Ascent Research.
