The ALG1L2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to produce a loss-of-function model for the ALG1L2 gene. Derived from HEK293T cells, this product uses CRISPR/Cas9-mediated gene disruption, yielding a mixed cellular pool that enables robust studies without single-cell cloning, reducing clonal artifacts in N-glycosylation research. The polyclonal format captures a spectrum of knockout events, providing a more physiologically representative system for functional genomics.
HEK293T cells are human embryonic kidney epithelial cells transformed with adenovirus and stably expressing SV40 large T antigen, which promotes episomal replication of SV40 origin-containing plasmids. These cells are highly transfectable and proliferate rapidly, making them a workhorse for recombinant protein expression and viral production. Their well-characterized genome and ease of manipulation render them an ideal chassis for studying complex post-translational modifications like glycosylation.
ALG1L2 encodes a putative alpha-1,3-mannosyltransferase that mediates early steps of dolichol-linked oligosaccharide biosynthesis in the ER, a prerequisite for N-linked glycosylation. It functions downstream of ER stress sensors ATF6, IRE1, and PERK and upstream of N-glycoprotein maturation, influencing receptor function and protein folding. ALG1L2 interacts with ALG13/ALG14 and DPM1, and participates in a cascade with ALG1, ALG2, and ALG11. Knockout may cause aberrant glycosylation and ER stress.
In the HEK293T cellular environment, ALG1L2 disruption offers a tractable system to dissect the immediate effects of impaired N-glycosylation. The model is particularly relevant for probing how defects in dolichol-linked oligosaccharide biosynthesis contribute to congenital disorders of glycosylation and the aberrant glycosylation patterns seen in cancer. Without clonal selection, the polyclonal population reflects diverse editing outcomes, allowing researchers to capture a range of phenotypic severities and analyze heterogeneous responses, including ER stress induction and altered glycoprotein trafficking.
This product facilitates detailed investigation of N-glycosylation pathway dynamics, functional comparison of ALG1L2 with its paralog ALG1, and dissection of ER stress signaling mechanisms. Researchers can employ a variety of analytical techniques, including western blotting for glycoprotein detection, lectin blotting to probe glycan structures, RT-qPCR for transcriptional profiling of glycosylation genes, ER stress reporter assays to monitor UPR activation, metabolic labeling with [^3H]mannose to assess sugar incorporation, and mass spectrometry-based glycomics for in-depth glycan analysis. For further information or to discuss customized applications, please contact Ascent Research.
