The RLIG1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the RLIG1 gene has been disrupted. This pooled knockout model provides a loss-of-function system for investigating RNA ligation and its associated pathways without clonal isolation, maintaining the inherent diversity of the edit pool.
HT29 is a human colorectal adenocarcinoma cell line with epithelial morphology, originally derived from a primary tumor. It is extensively used to study intestinal epithelial biology and colorectal cancer, retaining characteristics of enterocytic differentiation and tumorigenic signaling. The line??s epithelial nature makes it particularly suitable for examining RNA metabolism in the context of intestinal cell homeostasis and stress responses.
RLIG1 encodes an RNA ligase that joins RNA strands, playing essential roles in RNA repair and the final step of tRNA splicing. Together with the catalytic subunit RTCB, it forms part of the tRNA ligase complex. In the unfolded protein response (UPR), RLIG1 ligates XBP1 mRNA fragments generated by IRE1??, generating the spliced XBP1 transcription factor. Its expression is induced by ER stress sensors and HSF1, and its products include mature tRNA halves and XBP1s. Thus, RLIG1 connects RNA repair, tRNA processing, and the IRE1???CXBP1 UPR pathway.
Disruption of RLIG1 in HT29 polyclonal cells blocks RNA ligase function, leading to defective tRNA splicing and impaired XBP1s production. This abrogates the adaptive UPR, sensitizing the colorectal adenocarcinoma cells to ER stress and undermining pro-survival signals that support tumor cell growth. Consequently, the model permits precise interrogation of how RNA ligation-dependent stress responses influence colorectal cancer cell fitness and drug sensitivity.
This product enables studies of RNA repair in cancer, UPR signaling in colorectal adenocarcinoma, and identification of RLIG1 substrates via RNA immunoprecipitation and northern blotting. Functional assays include cell viability and colony formation under ER stress, RT-qPCR for XBP1 splicing, and western blotting for UPR markers. The model also supports drug screening targeting RNA metabolism or the UPR. For additional product information, please contact Ascent Research.
