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.
BLM hydrolase CRISPR/Cas9 KO Plasmid (h): sc-404411
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- BLM hydrolase 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 BLM hydrolase 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: BLM hydrolase Antibody (F-9): sc-166777
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
BLM hydrolase CRISPR/Cas9 KO Plasmid (h) | sc-404411 | 20 µg | $397.00 |
Overview
Human BLMH encodes bleomycin hydrolase, a cytosolic cysteine aminopeptidase involved in proteolytic processing of intracellular peptides and maintenance of protein quality control. BLMH participates in amino acid recycling and has been linked to cellular stress responses through effects on peptide turnover and proteostasis networks. Altered BLMH activity has been studied in contexts of drug metabolism and susceptibility to bleomycin-induced cytotoxicity, and has also been explored for potential connections to neurodegenerative and inflammatory phenotypes where peptide clearance pathways are perturbed. As a broadly expressed enzyme, BLMH provides a useful node for investigating how peptidase-dependent homeostasis intersects with oxidative stress and damage response signaling.
BLM hydrolase CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the BLMH gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the BLMH 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 BLMH 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 BLM hydrolase protein expression.
This CRISPR knockout system enables efficient generation of BLMH-deficient cell models for investigation of BLM hydrolase signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting BLMH exon(s) critical for BLM hydrolase 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 BLMH genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by BLM hydrolase CRISPR/Cas9 KO Plasmid (h) and BLM hydrolase CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the BLMH locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by BLM hydrolase HDR Plasmid (h) and BLM hydrolase HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by BLMH homology arms to support homology-directed repair at defined BLMH 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.