The KIAA0319L Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human KIAA0319L gene in the HAP1 cell line. This product provides a genetically disrupted cell pool in which the AAV receptor is ablated, enabling loss-of-function studies of KIAA0319L without clonal isolation. The polyclonal format retains population-level heterogeneity while ensuring robust gene disruption across the culture, offering a convenient and reproducible model for investigating AAV entry mechanisms and endocytic trafficking.
The parental HAP1 cell line is a near-haploid human cell line derived from the chronic myeloid leukemia (CML) line KBM-7, adapted to adherent growth. Its near-haploid karyotype simplifies genetic manipulation and facilitates functional genomics, as only one allele requires disruption for complete loss of function. HAP1 cells are widely employed in CRISPR-based screens and knockout studies due to their genetic tractability and stable growth characteristics. The adherent phenotype enables straightforward handling in transfection, transduction, and live-cell imaging experiments.
KIAA0319L encodes a type I transmembrane protein that functions as the critical receptor for adeno-associated virus (AAV), mediating viral entry via clathrin-mediated endocytosis. The protein interacts directly with AAV capsid proteins and recruits the clathrin adaptor complex AP2, initiating the formation of clathrin-coated pits. Subsequent internalization requires dynamin-dependent membrane scission, after which the virus?Creceptor complex is delivered to early endosomes marked by the early endosome antigen 1 (EEA1). Efficient AAV transduction in many cell types is strictly dependent on KIAA0319L, and its disruption abolishes viral infection. This polyclonal knockout population provides a clean background to dissect the AAV entry pathway and explore novel functions of this receptor.
In HAP1 cells, knockout of KIAA0319L eliminates AAV transduction, providing a powerful isogenic model to examine the absolute requirement for this receptor in viral entry. The near-haploid nature of HAP1 cells ensures a homogeneous loss-of-function phenotype upon gene disruption, minimizing the confounding effects of heterozygosity. Consequently, these polyclonal knockout cells are ideally suited for comparing wild-type and knockout responses to AAV serotypes, identifying alternative receptors, or performing genetic screens to uncover host factors involved in endocytosis. This cell model also serves as a stringent control for AAV-based gene delivery experiments, validating on-target effects of engineered vectors.
These polyclonal knockout cells enable a wide range of experimental applications, including detailed mechanistic studies of AAV internalization using transduction assays with reporter vectors (e.g., luciferase or GFP), co-immunoprecipitation of viral capsid proteins, and immunofluorescence-based tracking of viral particles. They are also valuable for investigating clathrin-mediated endocytosis dynamics through inhibitor studies and for functional genomics screens to identify modulators of viral entry. Additionally, the cells serve as a negative control in AAV vector development and gene therapy research. For further information on this product, please contact Ascent Research.
