The CD46 Knockout HAP1 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population, designed to disrupt the CD46 gene in the HAP1 human cell line. This polyclonal pool provides a heterogeneous knockout model suitable for loss-of-function studies without single-cell cloning, enabling robust analysis of CD46-dependent processes. The targeted gene disruption abrogates endogenous CD46 expression, offering a versatile tool for investigating its functions in complement regulation, viral pathogenesis, and immune signaling.
The HAP1 cell line is a near-haploid human cell line originally isolated from a patient with chronic myeloid leukemia. Its haploid karyotype facilitates genetic knockout and screening applications, as only one allele requires editing to achieve functional gene disruption. HAP1 cells retain many characteristics of their myeloid leukemia origin while providing a simplified genetic background, making them an advantageous host for CRISPR-based functional genomics and pathway dissection.
CD46, also known as membrane cofactor protein (MCP), serves as a critical complement regulatory protein by binding opsonins C3b and C4b and acting as a cofactor for their proteolytic inactivation by factor I, thereby protecting host cells from complement-mediated lysis. Beyond complement, CD46 functions as a cellular receptor for measles virus hemagglutinin and adenovirus fiber, mediating viral entry. Upon engagement, CD46 triggers intracellular signaling cascades: it interacts with GOPC to promote autophagy via VPS34 and LC3, modulates T-cell responses through AKT signaling and IL-10 production, and suppresses IFN-??. These pathways are orchestrated in concert with interacting partners such as CD9 and DLG1, linking innate immunity, infection, and adaptive immune regulation.
In the HAP1 background, CD46 knockout removes the cell’s intrinsic complement regulatory shield and disrupts viral entry receptors, creating a loss-of-function model to parse CD46’s dual roles. The haploid genetic context amplifies phenotypic penetrance, enabling clear elucidation of CD46-dependent mechanisms in resistance to complement lysis and susceptibility to measles virus or adenovirus infection. This system is particularly valuable for studying how CD46 coordinates autophagy induction and T-cell modulation, as HAP1 cells express relevant downstream effectors like GOPC, VPS34, and AKT.
These polyclonal knockout cells are suited for diverse research applications including functional genomics screens, complement regulation assays (e.g., C3b/C4b cleavage and lysis protection), viral infection studies with measles or adenovirus, autophagy flux analyses, T-cell activation assays, and co-immunoprecipitation of CD46 interactors. They support drug target validation in atypical hemolytic uremic syndrome, multiple sclerosis, and cancer, where CD46 is implicated. For further details, please contact Ascent Research.
