Description

The APOBEC1 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited human cell product in which the APOBEC1 gene has been functionally disrupted. Derived from the HEK293T host, this knockout cell line provides a stable loss-of-function model for studying RNA editing, lipoprotein metabolism, and post-transcriptional gene regulation. This cell line is supplied as a live product for immediate use in advanced research applications. The CRISPR/Cas9-mediated gene disruption abrogates APOBEC1 function, offering a robust alternative to transient knockdown methods.

The HEK293T parental cell line originates from human embryonic kidney epithelium and has been immortalized through stable integration of the SV40 large T antigen. This genetic modification enables episomal replication of transfected plasmids containing an SV40 origin of replication, leading to high-level expression of recombinant proteins and efficient production of lentiviral and retroviral vectors. Consequently, HEK293T cells are a cornerstone in molecular biology and drug discovery efforts, offering easy transfection, rapid growth, and scalability. Their human epithelial origin makes them a relevant model for studying intracellular signaling and metabolism, particularly when combined with targeted gene disruption.

APOBEC1 encodes the catalytic subunit of the RNA editing complex that deaminates cytidine 6666 to uridine in ApoB mRNA, producing the ApoB48 isoform essential for chylomicron assembly. The editing requires the cofactor ACF and involves interactions with RBM47, CUGBP2, hnRNP A/B, and KHSRP. Regulatory inputs include insulin, fatty acids, bile acids, HNF4??, and PPAR??; downstream targets comprise ApoB48, NAT1, and NF1. APOBEC1 also participates in innate antiviral responses.

In HEK293T cells, APOBEC1 knockout creates a simplified system for dissecting RNA editing mechanisms independent of tissue-specific factors. Leveraging the scalable, transfection-friendly nature of these cells, the model allows biochemical reconstitution and structure-function analysis of the editing complex. The line is ideal for substrate specificity studies, investigation of editing site recognition, and rescue experiments with APOBEC1 variants.

This cell line supports diverse assays: RNA editing analysis by RT-PCR and sequencing, co-immunoprecipitation of editing complex components (e.g., ACF), RNA immunoprecipitation, and immunofluorescence microscopy. It is applicable to lipid metabolism research, drug target validation, and antiviral studies. These applications are supported by well-established protocols for each assay type. For further information, contact Ascent Research.