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.
CALM CRISPR/Cas9 KO Plasmid (m): sc-433276
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CALM CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 CALM 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: CALM Antibody (A-2): sc-271224
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CALM CRISPR/Cas9 KO Plasmid (m) | sc-433276 | 20 µg | $397.00 |
Overview
Picalm (phosphatidylinositol binding clathrin assembly protein) encodes CALM, a cytosolic adaptor that couples phosphoinositide binding to clathrin-coated pit formation and cargo selection during endocytosis. CALM regulates receptor internalization, synaptic vesicle recycling, and endosome trafficking, thereby influencing membrane homeostasis and downstream signaling dynamics. In mouse systems, perturbation of Picalm can alter intracellular transport routes and autophagy-lysosome flux, processes often examined in neuronal and hematopoietic contexts. Genetic and functional studies have linked PICALM/CALM biology to neurodegeneration-related trafficking defects and to dysregulated cell growth programs through altered receptor turnover and vesicular transport.
CALM CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Picalm gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Picalm 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 Picalm 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 CALM protein expression.
This CRISPR knockout system enables efficient generation of Picalm-deficient cell models for investigation of CALM signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Picalm exon(s) critical for CALM 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 Picalm genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by CALM CRISPR/Cas9 KO Plasmid (m) and CALM CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Picalm locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by CALM HDR Plasmid (m) and CALM HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Picalm homology arms to support homology-directed repair at defined Picalm 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.