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.
ITM1 CRISPR/Cas9 KO Plasmid (h): sc-405155
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ITM1 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 ITM1 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: ITM1 Antibody (A-2): sc-390227
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ITM1 CRISPR/Cas9 KO Plasmid (h) | sc-405155 | 20 µg | $397.00 |
Overview
Human STT3A encodes the catalytic subunit of the oligosaccharyltransferase (OST) complex that mediates co‑translational N‑linked glycosylation in the endoplasmic reticulum. By transferring preassembled oligosaccharides to asparagine residues within nascent polypeptides, STT3A supports protein folding, quality control, and trafficking, and influences ER-associated degradation and proteostasis. Perturbation of STT3A-dependent glycosylation can disrupt membrane and secreted protein maturation, alter receptor and immune signaling, and engage the unfolded protein response. Dysregulated N‑glycosylation pathways have been linked to congenital disorders of glycosylation and are frequently remodeled in cancer and other diseases characterized by ER stress and altered cell-surface glycoproteomes.
ITM1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the STT3A gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the STT3A 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 STT3A 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 ITM1 protein expression.
This CRISPR knockout system enables efficient generation of STT3A-deficient cell models for investigation of ITM1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting STT3A exon(s) critical for ITM1 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 STT3A genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ITM1 CRISPR/Cas9 KO Plasmid (h) and ITM1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the STT3A locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ITM1 HDR Plasmid (h) and ITM1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by STT3A homology arms to support homology-directed repair at defined STT3A 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.