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.
SPRED1 CRISPR/Cas9 KO Plasmid (h): sc-402981
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SPRED1 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 SPRED1 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: SPRED1 Antibody (E-5): sc-393198
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SPRED1 CRISPR/Cas9 KO Plasmid (h) | sc-402981 | 20 µg | $397.00 |
Overview
SPRED1 (sprouty-related EVH1 domain-containing protein 1) is a negative regulator of receptor tyrosine kinase signaling that restrains the Ras–RAF–MEK–ERK/MAPK cascade by coupling to the neurofibromin (NF1) Ras-GAP complex. By limiting growth factor–dependent ERK activation, SPRED1 influences cell proliferation, differentiation, and migration across multiple tissue contexts. Loss-of-function variants in SPRED1 are associated with Legius syndrome and altered pigmentation and neurodevelopmental features, and reduced SPRED1 activity can impact MAPK pathway amplitude in cancer-relevant signaling networks. SPRED1 is therefore widely studied in models of Rasopathy biology, ERK-driven transcriptional programs, and feedback control of mitogenic signaling.
SPRED1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SPRED1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SPRED1 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 SPRED1 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 SPRED1 protein expression.
This CRISPR knockout system enables efficient generation of SPRED1-deficient cell models for investigation of SPRED1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SPRED1 exon(s) critical for SPRED1 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 SPRED1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by SPRED1 CRISPR/Cas9 KO Plasmid (h) and SPRED1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SPRED1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by SPRED1 HDR Plasmid (h) and SPRED1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SPRED1 homology arms to support homology-directed repair at defined SPRED1 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.