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.
Rho T1 CRISPR/Cas9 KO Plasmid (h): sc-401601
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rho T1 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 Rho T1 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: Rho T1 Antibody (A-8): sc-398520
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rho T1 CRISPR/Cas9 KO Plasmid (h) | sc-401601 | 20 µg | $397.00 |
Overview
RHOT1 encodes Rho T1 (Miro1), an atypical Rho-family GTPase anchored to the outer mitochondrial membrane that couples mitochondria to microtubule-based transport through interactions with TRAK adaptors and kinesin/dynein motors. By integrating GTPase activity with EF-hand calcium sensing, Rho T1 coordinates mitochondrial trafficking, positioning, and turnover, influencing neuronal polarization, synaptic function, and energy distribution. RHOT1 also contributes to mitochondrial quality control via crosstalk with PINK1/Parkin-dependent mitophagy pathways and regulation of organelle dynamics at ER–mitochondria contact sites. Dysregulation of RHOT1-dependent transport and mitophagy has been linked to cellular stress responses and is frequently studied in the context of neurodegeneration and other disorders characterized by mitochondrial dysfunction.
Rho T1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RHOT1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RHOT1 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 RHOT1 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 Rho T1 protein expression.
This CRISPR knockout system enables efficient generation of RHOT1-deficient cell models for investigation of Rho T1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting RHOT1 exon(s) critical for Rho T1 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 RHOT1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Rho T1 CRISPR/Cas9 KO Plasmid (h) and Rho T1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RHOT1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Rho T1 HDR Plasmid (h) and Rho T1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RHOT1 homology arms to support homology-directed repair at defined RHOT1 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.