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.
MARCKS CRISPR/Cas9 KO Plasmid (h): sc-400837
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MARCKS 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 MARCKS 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: MARCKS Antibody (JK-8): sc-100777
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MARCKS CRISPR/Cas9 KO Plasmid (h) | sc-400837 | 20 µg | $397.00 |
Overview
MARCKS (myristoylated alanine-rich C-kinase substrate) is a membrane-associated, actin-regulatory protein that functions as a prominent substrate of protein kinase C and modulates phosphatidylinositol 4,5-bisphosphate (PIP2) availability at the plasma membrane. Through phosphorylation-dependent cycling between membrane and cytoskeletal compartments, MARCKS influences actin filament organization, cell shape, adhesion, motility, vesicle trafficking, and neurite outgrowth. MARCKS participates in signaling networks linked to PKC, calcium/calmodulin regulation, and phosphoinositide dynamics that coordinate cytoskeletal remodeling and membrane signaling. Dysregulated MARCKS expression or phosphorylation has been associated with altered migration and invasion phenotypes and has been studied in contexts including cancer biology, neurodevelopmental processes, and inflammatory signaling.
MARCKS CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the MARCKS gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the MARCKS 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 MARCKS 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 MARCKS protein expression.
This CRISPR knockout system enables efficient generation of MARCKS-deficient cell models for investigation of MARCKS signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting MARCKS exon(s) critical for MARCKS 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 MARCKS genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MARCKS CRISPR/Cas9 KO Plasmid (h) and MARCKS CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the MARCKS locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MARCKS HDR Plasmid (h) and MARCKS HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by MARCKS homology arms to support homology-directed repair at defined MARCKS 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.