The ACADS Knockout A-549 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human cell line, designed to disrupt expression of the ACADS gene. This polyclonal mixture provides a loss-of-function model for studying the mitochondrial enzyme short-chain acyl-CoA dehydrogenase (SCAD), enabling dissection of short-chain fatty acid metabolism in a cancer-relevant background. Unlike clonal lines, this polyclonal pool captures the heterogeneity of gene disruption across a cell population, suitable for pooled functional assays.
The A-549 host cell line originates from a 58-year-old Caucasian male with lung carcinoma and is widely employed as a model of alveolar basal epithelial cells. A-549 cells exhibit an adherent epithelial morphology and are extensively used in lung adenocarcinoma research, particularly for studies of cancer signaling, drug response, and metabolic reprogramming. Their robust growth and well-characterized genetic background make them a reliable platform for knockout modeling.
ACADS encodes the mitochondrial matrix enzyme short-chain acyl-CoA dehydrogenase, which catalyzes the ??,??-dehydrogenation of short-chain (C4-C6) acyl-CoA esters, initiating the first step of the ??-oxidation spiral for these substrates. The reaction requires FAD as a cofactor and transfers electrons to electron transfer flavoprotein (ETF). ACADS expression is transcriptionally upregulated by the nuclear receptor PPARA and its coactivator PPARGC1A in response to fasting and glucagon signaling. The resulting enoyl-CoA product is further processed through sequential activities of enoyl-CoA hydratase (ECHS1) and ??-hydroxyacyl-CoA dehydrogenase (HADH) to generate acetyl-CoA, which feeds the tricarboxylic acid cycle or ketone body synthesis; acetyl-CoA acetyltransferase 1 (ACAT1) is a downstream enzyme in ketone body utilization pathways.
Disruption of ACADS in A-549 cells is particularly relevant for investigating the intersection of fatty acid oxidation and cancer metabolism. Lung adenocarcinoma cells often exhibit metabolic plasticity, and compromised short-chain fatty acid oxidation may sensitize them to nutrient stress or alter their proliferative capacity. This knockout model thus enables examination of ACADS-dependent metabolic contributions in a carcinoma context, with potential implications for understanding lipid metabolism disorders such as short-chain acyl-CoA dehydrogenase deficiency.
Researchers can employ this polyclonal knockout pool in a variety of functional assays, including Western blot and qRT-PCR to confirm ACADS disruption, fatty acid oxidation measurements using labeled substrates, metabolic flux analysis to track carbon routing, and cell viability assessments under glucose starvation to probe metabolic vulnerabilities. This model supports studies of short-chain fatty acid metabolism, metabolic disorder modeling, and cancer metabolism research, offering a versatile tool for interrogating ACADS function. For additional technical details and inquiry, please contact Ascent Research.
