GRAMD1B CRISPR/Cas9 KO Plasmid (h): sc-408929
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GRAMD1B 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 GRAMD1B 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 | |
GRAMD1B CRISPR/Cas9 KO Plasmid (h) | sc-408929 | 20 µg | $397.00 |
Overview
GRAMD1B (GRAM domain-containing 1B) is an ER-anchored cholesterol transfer protein that participates in nonvesicular lipid transport at endoplasmic reticulum–plasma membrane contact sites. By binding accessible cholesterol via its StART-like domain and responding to phosphoinositide cues at the plasma membrane, GRAMD1B contributes to cholesterol homeostasis, membrane organization, and lipid-dependent signaling. These processes intersect with sterol sensing, ER stress responses, and broader metabolic regulation that influence cell growth and survival programs. Altered GRAMD1B expression or activity has been investigated in contexts of dysregulated lipid metabolism and cancer-associated signaling, making it relevant for mechanistic studies of membrane biology.
GRAMD1B CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the GRAMD1B gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the GRAMD1B 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 GRAMD1B 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 GRAMD1B protein expression.
This CRISPR knockout system enables efficient generation of GRAMD1B-deficient cell models for investigation of GRAMD1B signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting GRAMD1B exon(s) critical for GRAMD1B 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 GRAMD1B genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by GRAMD1B CRISPR/Cas9 KO Plasmid (h) and GRAMD1B CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the GRAMD1B locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by GRAMD1B HDR Plasmid (h) and GRAMD1B HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by GRAMD1B homology arms to support homology-directed repair at defined GRAMD1B 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.