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.
Amylase 2a CRISPR/Cas9 KO Plasmid (h): sc-400533
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Amylase 2a 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 Amylase 2a 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Amylase 2a CRISPR/Cas9 KO Plasmid (h) | sc-400533 | 20 µg | $397.00 |
Overview
AMY2A encodes pancreatic α-amylase 2A, a secreted hydrolase that initiates dietary starch and glycogen breakdown by cleaving α-1,4-glycosidic bonds to generate maltose and oligosaccharides. As part of the exocrine pancreatic enzyme program, Amylase 2a contributes to digestive metabolism and is regulated within acinar cell differentiation and secretory pathway processes, including ER/Golgi trafficking and stimulus-coupled enzyme release. Altered amylase expression or activity is used as a molecular readout of pancreatic exocrine function and is frequently examined in contexts of pancreatic inflammation and pancreatic disorders where acinar cell homeostasis is perturbed. AMY2A also serves as a marker in studies of acinar cell identity, stress responses, and digestive enzyme gene regulation.
Amylase 2a CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the AMY2A gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the AMY2A 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 AMY2A 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 Amylase 2a protein expression.
This CRISPR knockout system enables efficient generation of AMY2A-deficient cell models for investigation of Amylase 2a signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting AMY2A exon(s) critical for Amylase 2a 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 AMY2A genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Amylase 2a CRISPR/Cas9 KO Plasmid (h) and Amylase 2a CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the AMY2A locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Amylase 2a HDR Plasmid (h) and Amylase 2a HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by AMY2A homology arms to support homology-directed repair at defined AMY2A 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.