The EFEMP1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated by targeted disruption of the EFEMP1 gene. This loss-of-function model is produced in the HAP1 human near-haploid cell line, yielding a genetically diverse pool of knockout cells suitable for functional studies. The gene product, fibulin-3, is an extracellular matrix glycoprotein implicated in cell adhesion, migration, and tissue organization. This product enables researchers to explore EFEMP1??s roles in matrix biology, signal transduction, and disease-associated pathways without clonal selection artifacts.
HAP1 cells are a near-haploid, fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia line. Their functional haploidy simplifies gene editing and phenotypic interpretation, making them a widely used model for CRISPR-based knockout screens and mechanistic studies. HAP1 cells retain active adhesion and migration pathways, thus providing a sensitive background for analyzing extracellular matrix and integrin-mediated signaling. The knockout of EFEMP1 in this host enables clean assessment of fibulin-3 functions in a cell model with minimal genetic redundancy.
EFEMP1 encodes fibulin-3, an extracellular matrix glycoprotein that directly binds collagen type I alpha 2 (COL1A2), fibronectin (FN1), integrin beta-1 (ITGB1), and transforming growth factor beta 1 (TGFB1). Its expression is regulated by TGFB1 and epidermal growth factor (EGF), and it modulates downstream matrix metalloproteinases MMP2 and MMP9, integrin-dependent adhesion, and cell cycle progression. Through ITGB1 engagement, fibulin-3 activates focal adhesion kinase (FAK) and SRC kinases, leading to MAPK1/MAPK3 (ERK2/ERK1) phosphorylation and cytoskeletal reorganization. Concurrent modulation of TGF-beta and Wnt signaling further influences matrix remodeling and cellular responses.
In the HAP1 background, disruption of EFEMP1 perturbs cell?Cmatrix interactions, providing a tractable system to dissect the molecular basis of fibulin-3-related pathologies such as Doyne honeycomb retinal dystrophy and age-related macular degeneration. The near-haploid nature minimizes genetic buffering, ensuring that phenotypic changes are directly attributable to EFEMP1 loss. This model is also valuable for cancer biology, as fibulin-3 has been implicated in tumor cell invasion and metastatic dissemination through matrix remodeling. Consequently, the knockout cells serve as a robust platform for elucidating EFEMP1’s contributions to both ocular and oncological disorders.
These knockout cells enable detailed studies of extracellular matrix biology, adhesion signaling, and disease modeling. Typical readouts include western blotting, RT?qPCR, and immunofluorescence for pathway activation, alongside cell migration and adhesion assays to quantify phenotypic changes. Co?immunoprecipitation facilitates interrogation of altered interactions with partners such as ITGB1, FN1, and TGFB1. Applications span from cancer metastasis, where fibulin?3 influences invasion, to retinal degeneration research addressing matrix dysregulation in macular disease. For additional information, please contact Ascent Research.
