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.
hamartin CRISPR/Cas9 KO Plasmid (h): sc-402444
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- hamartin 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 hamartin 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: hamartin Antibody (C-8): sc-377386
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
hamartin CRISPR/Cas9 KO Plasmid (h) | sc-402444 | 20 µg | $397.00 |
Overview
TSC1 encodes hamartin, a scaffold protein that forms a functional complex with TSC2 (tuberin) to restrain mTORC1 signaling by regulating the small GTPase Rheb. Through this tumor suppressor axis, hamartin integrates inputs from PI3K–AKT, AMPK, and cellular stress cues to coordinate protein synthesis, autophagy, metabolism, and cell growth. Loss or dysfunction of TSC1 disrupts mTOR-dependent homeostasis, altering lysosomal and cytoskeletal dynamics and perturbing neuronal and glial development. These pathways are central to mechanisms studied in tuberous sclerosis complex biology and broader mTOR-driven cell growth and differentiation phenotypes.
hamartin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the TSC1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the TSC1 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 TSC1 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 hamartin protein expression.
This CRISPR knockout system enables efficient generation of TSC1-deficient cell models for investigation of hamartin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting TSC1 exon(s) critical for hamartin 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 TSC1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by hamartin CRISPR/Cas9 KO Plasmid (h) and hamartin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the TSC1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by hamartin HDR Plasmid (h) and hamartin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by TSC1 homology arms to support homology-directed repair at defined TSC1 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.