The KIAA0319L Knockout HT29 Polyclonal Cells represent a polyclonal population of HT29 human colorectal adenocarcinoma cells in which the KIAA0319L gene, encoding the adeno-associated virus receptor (AAVR), has been disrupted by CRISPR/Cas9-mediated genome editing. This loss-of-function model is designed for research into AAV transduction mechanisms and host?Cpathogen interactions.
The HT29 host cell line originates from a colorectal adenocarcinoma of a 44-year-old Caucasian female. It harbors mutations in APC and TP53 and can be induced to differentiate with sodium butyrate, providing a clinically relevant intestinal epithelial model for investigating colorectal cancer, epithelial barrier function, and differentiation. This genetic context is particularly valuable for studying how viral entry pathways intersect with oncogenic signaling.
KIAA0319L serves as the primary receptor for AAV, facilitating viral attachment and clathrin-mediated endocytosis. The receptor interacts with AAV capsid proteins and recruits the AP2 adaptor complex, clathrin heavy chain (CLTC), and dynamin-2 (DNM2) to drive vesicle internalization. Internalized virus then traffics through early endosomes positive for RAB5 and EEA1, maturing into late endosomes/lysosomes marked by RAB7 and LAMP1; the NPC1 cholesterol transporter also contributes to capsid processing. Upstream transcriptional control of KIAA0319L is not well-documented, but may respond to growth factor stimulation and oncogenic signaling pathways active in HT29 cells.
In the context of HT29 cells, knockout of KIAA0319L abolishes AAV transduction, creating a null background for dissecting viral entry requirements and receptor function. This model permits exploration of potential KIAA0319L roles in cell adhesion or neurodevelopment, as suggested by genetic associations with reading disabilities. Additionally, the colorectal cancer setting allows investigation of any interplay between AAV trafficking and malignancy-related pathways.
Research applications include AAV transduction assays with luciferase or GFP reporters, flow cytometry to quantify viral uptake, and immunofluorescence monitoring of capsid trafficking. The model supports co-immunoprecipitation of AAVR with viral particles, endocytosis studies with labeled transferrin, and CRISPR validation by Sanger sequencing. It is highly valuable for gene therapy vector engineering and host-pathogen interaction research in a colorectal cancer background. Cell viability assays ensure the knockout does not impair general cellular functions. For further information, please contact Ascent Research.
