SPEN CRISPR/Cas9 KO Plasmid (h): sc-406083
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SPEN 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 SPEN 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 | |
SPEN CRISPR/Cas9 KO Plasmid (h) | sc-406083 | 20 µg | $397.00 |
Overview
SPEN (also known as SHARP) encodes a large nuclear RNA-binding transcriptional coregulator that couples sequence-specific DNA-binding partners and long noncoding RNAs to chromatin-modifying complexes. Through interactions with SMRT/HDAC and related repressors, SPEN contributes to transcriptional silencing programs, including XIST-dependent X-chromosome inactivation, and influences lineage-specific gene expression. SPEN also modulates signaling-responsive transcriptional outputs, with reported connections to Notch-associated regulation and broader epigenetic control of developmental and immune-related pathways. Dysregulated SPEN function or expression has been implicated in altered differentiation states and transcriptional rewiring observed across multiple disease contexts, supporting its utility as a mechanistic node in gene regulation studies.
SPEN CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SPEN gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SPEN 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 SPEN 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 SPEN protein expression.
This CRISPR knockout system enables efficient generation of SPEN-deficient cell models for investigation of SPEN signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SPEN exon(s) critical for SPEN 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 SPEN genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by SPEN CRISPR/Cas9 KO Plasmid (h) and SPEN CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SPEN locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by SPEN HDR Plasmid (h) and SPEN HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SPEN homology arms to support homology-directed repair at defined SPEN 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.