Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
SAHH CRISPR/Cas9 KO Plasmid (h): sc-402617
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SAHH CRISPR/Cas9 Knockout (KO) Plasmid (h) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the SAHH genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: SAHH Antibody (A-11): sc-271389
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SAHH CRISPR/Cas9 KO Plasmid (h) | sc-402617 | 20 µg | $397.00 |
Overview
AHCY encodes S-adenosylhomocysteine hydrolase (SAHH), a key enzyme in the methionine cycle that hydrolyzes S-adenosylhomocysteine to adenosine and homocysteine, thereby sustaining cellular methylation capacity by relieving feedback inhibition of S-adenosylmethionine-dependent methyltransferases. Through this role, SAHH influences epigenetic regulation, RNA and protein methylation, and phospholipid biosynthesis, integrating one-carbon metabolism with broader control of gene expression programs. Perturbation of AHCY activity can shift methylation potential and metabolic homeostasis, affecting pathways linked to proliferation, differentiation, and stress responses. Altered SAHH function has been studied in the context of methylation-associated disorders and cellular phenotypes relevant to cancer biology and immune and neurological research models.
SAHH CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the AHCY gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the AHCY 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 AHCY 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 SAHH protein expression.
This CRISPR knockout system enables efficient generation of AHCY-deficient cell models for investigation of SAHH signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting AHCY exon(s) critical for SAHH 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 AHCY genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by SAHH CRISPR/Cas9 KO Plasmid (h) and SAHH CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the AHCY locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by SAHH HDR Plasmid (h) and SAHH HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by AHCY homology arms to support homology-directed repair at defined AHCY 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.