Aldehyde dehydrogenase 5-A1/SSADH/ALDH5A1 CRISPR/Cas9 KO Plasmid (h): sc-406757

Overview

ALDH5A1 encodes succinic semialdehyde dehydrogenase (SSADH), a mitochondrial NAD+-dependent enzyme that catalyzes oxidation of succinic semialdehyde to succinate, linking GABA catabolism to the tricarboxylic acid (TCA) cycle. By controlling flux through the GABA shunt, ALDH5A1 helps regulate cellular redox balance, mitochondrial metabolism, and neurotransmitter turnover in neural tissues. Disruption of SSADH activity is associated with accumulation of GABA-derived metabolites and altered inhibitory signaling, and ALDH5A1 has been studied in the context of inborn errors of metabolism affecting the nervous system. These features make ALDH5A1 a useful target for probing metabolic crosstalk between mitochondrial function and neurotransmitter pathways.

Aldehyde dehydrogenase 5-A1/SSADH/ALDH5A1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ALDH5A1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ALDH5A1 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 ALDH5A1 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 5-A1/SSADH/ALDH5A1 protein expression.

This CRISPR knockout system enables efficient generation of ALDH5A1-deficient cell models for investigation of Aldehyde dehydrogenase 5-A1/SSADH/ALDH5A1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting ALDH5A1 exon(s) critical for Aldehyde dehydrogenase 5-A1/SSADH/ALDH5A1 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 ALDH5A1 genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by Aldehyde dehydrogenase 5-A1/SSADH/ALDH5A1 CRISPR/Cas9 KO Plasmid (h) and Aldehyde dehydrogenase 5-A1/SSADH/ALDH5A1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ALDH5A1 locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by Aldehyde dehydrogenase 5-A1/SSADH/ALDH5A1 HDR Plasmid (h) and Aldehyde dehydrogenase 5-A1/SSADH/ALDH5A1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ALDH5A1 homology arms to support homology-directed repair at defined ALDH5A1 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.