TBC1D24 CRISPR/Cas9 KO Plasmid (m): sc-432462
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TBC1D24 CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 TBC1D24 genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- TBC1D24 HDR Plasmid (m) (sc-432462-HDR) is recommended for co-transfection with TBC1D24 CRISPR/Cas9 KO Plasmid (m) to enable selection of successfully edited cells through HDR-mediated integration of a puromycin resistance cassette and RFP reporter gene
- TBC1D24 HDR Plasmid (m) is a pool of plasmids, each containing a homology-directed repair (HDR) template corresponding to the gRNA target sites in the TBC1D24 CRISPR/Cas9 KO Plasmid (m)
- Each HDR plasmid contains two ~800 bp homology arms flanking the puromycin resistance and RFP cassettes, designed to bind genomic DNA sequences surrounding the Cas9-induced double-strand break site and facilitate precise HDR-mediated integration
- 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: TBC1D24 Antibody (G-6): sc-390237
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TBC1D24 CRISPR/Cas9 KO Plasmid (m) | sc-432462 | 20 µg | $397.00 | |||
TBC1D24 HDR Plasmid (m) | sc-432462-HDR | 20 µg | $445.00 |
Overview
Tbc1d24 encodes TBC1D24, a Tre2/Bub2/Cdc16 (TBC) domain–containing protein implicated in regulation of membrane trafficking and synaptic vesicle dynamics in neurons. Through interactions that influence Rab GTPase-associated processes and cytoskeletal organization, TBC1D24 contributes to endocytic recycling, vesicle turnover, and maintenance of neuronal excitability. Disruption of TBC1D24 function has been linked to neurodevelopmental and seizure-related phenotypes, making it relevant for studies of synaptic homeostasis and network stability in the mammalian brain. In mouse model systems, Tbc1d24 perturbation supports mechanistic investigation of neuronal maturation, vesicle trafficking pathways, and stress responses that shape circuit function.
TBC1D24 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tbc1d24 gene in mouse cell lines. Each plasmid in the pool co-expresses a unique sgRNA, targeting a distinct site within the Tbc1d24 locus, alongside the Streptococcus pyogenes Cas9 nuclease, and encodes GFP to enable fluorescent identification and enrichment of successfully transfected cells. This multi-guide strategy increases the likelihood of inducing frameshifts or deletions that produce a functional knockout, offering a more robust alternative to single-guide approaches. DSBs induced at multiple sites are resolved through non-homologous end joining (NHEJ) or, when used with the included HDR donor template, homology-directed repair (HDR) at a defined target site within the locus.
When used in conjunction with the RFP-expressing HDR donor, GFP and RFP fluorescence can be used together to distinguish transfected from edited cell populations, streamlining flow cytometry-based sorting and clone selection workflows.
Homology-Directed Repair (HDR) Donor — Puromycin Cassette with RFP Reporter
For applications requiring confirmed, selectable knockout clones, TBC1D24 HDR Plasmid (m) includes an HDR donor construct containing a puromycin resistance cassette (PuroR) and a red fluorescent protein (RFP) reporter, flanked by homology arms specific to a defined Tbc1d24 target site.
When co-transfected with TBC1D24 CRISPR/Cas9 KO Plasmid (m):
- The PuroR-RFP cassette integrates at the Cas9 cut site via HDR, disrupting the Tbc1d24 open reading frame.
- RFP fluorescence provides an immediate visual indicator of successful integration, enabling fluorescence-based identification or sorting of edited cells prior to or alongside puromycin selection.
- Successfully edited cells are confirmed through puromycin resistance, substantially reducing clone screening burden.
- This selection strategy is ideal for generating stable, clonal KO cell lines for downstream functional studies, drug screening, or model development.
Cre-lox Cassette Removal System
The HDR donor construct features loxP sites flanking the PuroR-RFP selection cassette to allow clean marker removal following clone confirmation. Transient expression of Cre recombinase via the included Cre Vector: sc-418923 excises the cassette, leaving a minimal residual loxP site within the Tbc1d24 locus and eliminating potential confounding effects on downstream assays.
This two-step approach:
- Minimizes disruption to local chromatin architecture and neighboring regulatory elements
- Restores a near-native genomic context at the edited locus
- Enables reuse of the puromycin selection strategy in the same cell line for additional edits
Key Features
- gRNA targeting Tbc1d24 exon(s) critical for TBC1D24 function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- HDR donor with puromycin resistance for positive clone selection
- loxP-flanked PuroR cassette with Cre recombinase vector for seamless marker removal
- Supplied ready to use for delivery by transfection
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.