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.
HMCES CRISPR/Cas9 KO Plasmid (h): sc-405008
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HMCES 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 HMCES 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: HMCES Antibody (B-2): sc-514238
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HMCES CRISPR/Cas9 KO Plasmid (h) | sc-405008 | 20 µg | $397.00 |
Overview
HMCES (5-hydroxymethylcytosine binding, embryonic stem cell-specific) encodes a DNA-protective protein that functions at stalled replication forks and sites of base damage. HMCES forms a covalent DNA–protein crosslink intermediate that shields abasic (AP) sites and related lesions, helping coordinate replication-coupled repair and preserve genome stability. This activity intersects with DNA damage response signaling, fork protection, and pathways linked to mutagenesis control during S phase. Altered HMCES function is therefore relevant to studies of replication stress, genome instability phenotypes, and cancer-associated DNA repair dependencies.
HMCES CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the HMCES gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the HMCES 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 HMCES 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 HMCES protein expression.
This CRISPR knockout system enables efficient generation of HMCES-deficient cell models for investigation of HMCES signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting HMCES exon(s) critical for HMCES 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 HMCES genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by HMCES CRISPR/Cas9 KO Plasmid (h) and HMCES CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the HMCES locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by HMCES HDR Plasmid (h) and HMCES HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by HMCES homology arms to support homology-directed repair at defined HMCES 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.