Aldehyde dehydrogenase 6-A1/ALDH6A1 CRISPR/Cas9 KO Plasmid (h): sc-406553

Overview

Human ALDH6A1 encodes mitochondrial methylmalonate semialdehyde dehydrogenase, an NAD(P)+-dependent enzyme that oxidizes methylmalonate semialdehyde to propionyl-CoA during valine and pyrimidine catabolism. By feeding carbon into propionyl-CoA metabolism and downstream anaplerotic pathways, ALDH6A1 links branched-chain amino acid turnover to mitochondrial energy homeostasis and redox balance. Perturbation of ALDH6A1 activity can alter organic acid profiles and mitochondrial stress responses, making it relevant to studies of inborn errors of metabolism and broader mitochondrial dysfunction phenotypes. Its expression and function are also of interest in metabolic reprogramming contexts where amino acid catabolism supports proliferative or stress-adapted states.

Aldehyde dehydrogenase 6-A1/ALDH6A1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ALDH6A1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ALDH6A1 together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.

The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the ALDH6A1 open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish Aldehyde dehydrogenase 6-A1/ALDH6A1 protein expression.

This CRISPR knockout system enables efficient generation of ALDH6A1-deficient cell models for investigation of Aldehyde dehydrogenase 6-A1/ALDH6A1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting ALDH6A1 exon(s) critical for Aldehyde dehydrogenase 6-A1/ALDH6A1 function
• Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
• GFP reporter for identification of transfected cells
• Pool of plasmids targeting multiple ALDH6A1 genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by Aldehyde dehydrogenase 6-A1/ALDH6A1 CRISPR/Cas9 KO Plasmid (h) and Aldehyde dehydrogenase 6-A1/ALDH6A1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ALDH6A1 locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by Aldehyde dehydrogenase 6-A1/ALDH6A1 HDR Plasmid (h) and Aldehyde dehydrogenase 6-A1/ALDH6A1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ALDH6A1 homology arms to support homology-directed repair at defined ALDH6A1 target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.