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.
ALAS-E CRISPR/Cas9 KO Plasmid (h): sc-403752
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ALAS-E 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 ALAS-E 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: ALAS-E Antibody (D-4): sc-166739
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ALAS-E CRISPR/Cas9 KO Plasmid (h) | sc-403752 | 20 µg | $397.00 |
Overview
ALAS2 encodes the erythroid-specific 5-aminolevulinate synthase (ALAS-E), a mitochondrial, pyridoxal phosphate–dependent enzyme that catalyzes the first and rate-limiting step of heme biosynthesis by condensing glycine and succinyl‑CoA to form 5‑aminolevulinate. Its activity integrates mitochondrial metabolism with erythroid differentiation and hemoglobinization, supporting iron utilization and porphyrin production in developing red blood cells. ALAS-E is regulated by erythroid transcriptional programs and heme/iron availability, linking it to pathways controlling mitochondrial function and oxidative stress in erythroblasts. Genetic or functional perturbation of ALAS2 is associated with disordered porphyrin/heme synthesis and can be investigated in models of erythroid maturation and red cell pathophysiology.
ALAS-E CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ALAS2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ALAS2 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 ALAS2 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 ALAS-E protein expression.
This CRISPR knockout system enables efficient generation of ALAS2-deficient cell models for investigation of ALAS-E signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ALAS2 exon(s) critical for ALAS-E 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 ALAS2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ALAS-E CRISPR/Cas9 KO Plasmid (h) and ALAS-E CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ALAS2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ALAS-E HDR Plasmid (h) and ALAS-E HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ALAS2 homology arms to support homology-directed repair at defined ALAS2 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.