The NALF2 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited knockout cell line derived from human embryonic kidney HEK293T cells, providing a stable loss-of-function model for the NALF2 gene. NALF2 encodes a putative N-acetyltransferase, and its disruption enables investigation of protein acetylation dynamics. This cell line is supplied as a ready-to-use research tool, bypassing the variability of transient silencing methods. The CRISPR-mediated gene disruption ensures consistent, heritable knockout of the target locus, making it suitable for reproducible functional assays.
HEK293T is a widely utilized host cell line, originally derived from HEK293 cells by stable expression of the SV40 large T antigen. This modification permits episomal replication of plasmids containing the SV40 origin, significantly boosting transient protein expression and viral vector production. The cell line??s epithelial morphology, human origin, and high transfectability render it an ideal platform for studying human gene function. The NALF2 knockout variant retains all these advantageous properties, providing a robust cellular context for examining acetyltransferase biology.
The NALF2 protein is a putative N-acetyltransferase that catalyzes acetyl group transfer from acetyl-coenzyme A (acetyl-CoA) to lysine residues on substrate proteins. This post-translational modification influences protein stability, activity, localization, and interactions, forming a key node in lysine acetylation signaling. NALF2 activity is dependent on acetyl-CoA availability, which is governed by cellular metabolic state. Deacetylase enzymes, such as histone deacetylases (HDACs) and sirtuins (SIRTs), reverse this modification, creating a dynamic regulatory network. NALF2 thus links metabolic status to the acetyl-proteome.
In the HEK293T background, knockout of NALF2 allows unambiguous dissection of its role in acetylation-dependent signaling. The host cells possess active acetylation machinery and responsive metabolic pathways, making them physiologically relevant for mechanistic studies. By comparing the knockout line to wild-type controls, researchers can identify NALF2-specific substrates and downstream effects on processes such as signal transduction or metabolic regulation. The absence of NALF2 may reveal compensatory changes in deacetylase activity or alternative acetylation pathways.
This NALF2 knockout cell line supports a wide array of experimental applications, including functional characterization of acetyltransferase activity, acetyl-proteomics, and inhibitor screening. Representative assays include anti-acetyl-lysine western blotting to monitor global acetylation changes, RT-qPCR for verifying NALF2 knockout, in vitro acetyltransferase activity assays with recombinant NALF2, and co-immunoprecipitation to capture acetylated protein partners. LC-MS/MS acetyl-proteomics can comprehensively profile altered acetylation sites, while cell viability assays gauge functional outcomes. For technical details and customization, contact Ascent Research.
