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.
MBD3 CRISPR/Cas9 KO Plasmid (h): sc-401072
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MBD3 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 MBD3 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: MBD3 Antibody (F-11): sc-166319
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MBD3 CRISPR/Cas9 KO Plasmid (h) | sc-401072 | 20 µg | $397.00 |
Overview
MBD3 encodes methyl-CpG binding domain protein 3, a core component of the NuRD (nucleosome remodeling and deacetylase) complex that links ATP-dependent chromatin remodeling with histone deacetylation to regulate transcriptional programs. Unlike some MBD family members, MBD3 has limited affinity for methylated CpG DNA but is essential for targeting NuRD to specific genomic loci through protein–protein interactions and chromatin context. Through NuRD, MBD3 contributes to control of cell fate decisions, DNA damage responses, and maintenance of chromatin states that influence proliferation and differentiation. Dysregulation of MBD3/NuRD-associated epigenetic control has been implicated in aberrant transcriptional networks observed in cancer and in altered developmental gene regulation relevant to neurodevelopmental and stem cell biology studies.
MBD3 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the MBD3 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the MBD3 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 MBD3 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 MBD3 protein expression.
This CRISPR knockout system enables efficient generation of MBD3-deficient cell models for investigation of MBD3 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting MBD3 exon(s) critical for MBD3 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 MBD3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MBD3 CRISPR/Cas9 KO Plasmid (h) and MBD3 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the MBD3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MBD3 HDR Plasmid (h) and MBD3 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by MBD3 homology arms to support homology-directed repair at defined MBD3 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.