ACBD5 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with disruption of the ACBD5 gene. Derived from the HEK293T host, these cells provide a human cellular model for investigating peroxisomal very long-chain fatty acid (VLCFA) import and metabolism. The polyclonal format offers diverse editing outcomes suitable for population-level functional studies. This loss-of-function model enables research into ACBD5-dependent processes, including VLCFA transport and peroxisomal beta-oxidation.
The HEK293T cell line is a human embryonic kidney epithelial line stably expressing SV40 large T antigen, conferring high transfection efficiency and episomal plasmid replication. Widely used for transient protein expression and viral production, HEK293T cells contain endogenous peroxisomal machinery, making them ideal for mechanistic studies of peroxisome function. Their rapid growth and genetic tractability facilitate the generation of knockout models for biochemical and cell-based assays.
ACBD5 encodes a peroxisomal membrane protein that binds very long-chain fatty acyl-CoAs and facilitates their transfer to the ABCD1 transporter for peroxisomal import. This process is essential for beta-oxidation, with downstream activity of enzymes such as ACOX1, DBP, and SCP2. ACBD5 interacts with ABCD1 and peroxisomal biogenesis factors including PEX3, PEX16, and PEX19. Its expression is regulated by the PPAR??/RXR heterodimer, responsive to fibrates and fatty acid status. Knockout of ACBD5 disrupts VLCFA import, leading to metabolic accumulation and potential peroxisomal dysfunction.
In the HEK293T background, ACBD5 knockout provides a simplified platform to study VLCFA metabolism without confounding factors from primary tissues. The high transfection efficiency enables rapid rescue experiments and overexpression of pathway components, facilitating structure-function analyses. This model permits clear phenotypic assessment of peroxisomal import and beta-oxidation, with applications in lipidomic profiling and functional flux assays. It serves as a valuable tool for dissecting the molecular underpinnings of peroxisomal disorders.
Typical assays include immunofluorescence for peroxisomal markers, LC-MS/MS quantification of VLCFA levels, and beta-oxidation flux measurements using radiolabeled substrates. Co-immunoprecipitation can probe ACBD5-ABCD1 interactions, while RT-qPCR analyzes peroxisomal gene expression. Cell viability under fatty acid stress and, with differentiation, retinal cell phenotypes can be evaluated. This product supports research into peroxisomal biogenesis, retinal dystrophy, and X-linked adrenoleukodystrophy. For additional details, please contact Ascent Research.
