Tuba CRISPR/Cas9 KO Plasmid (h): sc-406569
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Tuba 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 Tuba 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: Tuba Antibody (MH-8): sc-135592
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Tuba CRISPR/Cas9 KO Plasmid (h) | sc-406569 | 20 µg | $397.00 |
Overview
DNMBP encodes Tuba, a multidomain Rho guanine nucleotide exchange factor that links membrane signaling to actin and microtubule cytoskeletal remodeling. Through activation of small GTPases such as CDC42 and coordination with endocytic and junctional complexes, Tuba helps regulate cell polarity, vesicle trafficking, and remodeling of adherens and tight junctions. These functions place DNMBP within pathways controlling epithelial organization, neurite outgrowth, and migration-dependent morphogenesis. Dysregulated cytoskeletal signaling and polarity networks involving Tuba have been implicated in contexts of aberrant cell invasion and neurodevelopmental phenotypes, supporting its value in mechanistic studies of disease-relevant cellular architecture.
Tuba CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the DNMBP gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the DNMBP 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 DNMBP 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 Tuba protein expression.
This CRISPR knockout system enables efficient generation of DNMBP-deficient cell models for investigation of Tuba signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting DNMBP exon(s) critical for Tuba 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 DNMBP genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Tuba CRISPR/Cas9 KO Plasmid (h) and Tuba CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the DNMBP locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Tuba HDR Plasmid (h) and Tuba HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by DNMBP homology arms to support homology-directed repair at defined DNMBP 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.