ALKBH4 CRISPR/Cas9 KO Plasmid (h): sc-406350
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ALKBH4 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 ALKBH4 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ALKBH4 CRISPR/Cas9 KO Plasmid (h) | sc-406350 | 20 µg | $397.00 |
Overview
ALKBH4 encodes a member of the AlkB family of Fe(II)/2-oxoglutarate–dependent dioxygenases implicated in oxidative demethylation reactions on nucleic acids and proteins. Human ALKBH4 has been linked to regulation of actin cytoskeleton dynamics and cellular architecture, consistent with roles in processes such as cell migration, adhesion, and division. Through these activities, ALKBH4 is often studied in the context of genome stability, stress responses, and transcriptional or post-transcriptional control. Altered ALKBH4 expression or function has been explored in multiple disease-relevant settings, including cancer-associated phenotypes where cytoskeletal remodeling and DNA/RNA metabolism are frequently dysregulated.
ALKBH4 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ALKBH4 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ALKBH4 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 ALKBH4 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 ALKBH4 protein expression.
This CRISPR knockout system enables efficient generation of ALKBH4-deficient cell models for investigation of ALKBH4 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ALKBH4 exon(s) critical for ALKBH4 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 ALKBH4 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ALKBH4 CRISPR/Cas9 KO Plasmid (h) and ALKBH4 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ALKBH4 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ALKBH4 HDR Plasmid (h) and ALKBH4 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ALKBH4 homology arms to support homology-directed repair at defined ALKBH4 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.