RyR-2 CRISPR/Cas9 KO Plasmid (h): sc-402986

Overview

RYR2 encodes ryanodine receptor 2 (RyR-2), a tetrameric intracellular Ca²⁺ release channel located on the sarcoplasmic/endoplasmic reticulum that mediates excitation–contraction coupling by generating calcium-induced calcium release. RyR-2 activity coordinates Ca²⁺ transients that regulate cardiomyocyte contraction, mitochondrial metabolism, and Ca²⁺-dependent signaling pathways including CaMKII- and calcineurin/NFAT-linked transcriptional responses. Channel function is shaped by macromolecular complex assembly, post-translational modification, and luminal Ca²⁺ sensing, integrating adrenergic inputs with intracellular Ca²⁺ homeostasis. Genetic and functional perturbations of RYR2 are associated with inherited arrhythmia syndromes and cardiomyopathy-related phenotypes, making it a central node for studying cardiac calcium handling and stress-responsive signaling.

RyR-2 CRISPR/Cas9 Knockout CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RYR2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RYR2 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 RYR2 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 RyR-2 CRISPR/Cas9 Knockout protein expression.

This CRISPR knockout system enables efficient generation of RYR2-deficient cell models for investigation of RyR-2 CRISPR/Cas9 Knockout signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting RYR2 exon(s) critical for RyR-2 CRISPR/Cas9 Knockout 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 RYR2 genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by RyR-2 CRISPR/Cas9 Knockout CRISPR/Cas9 KO Plasmid (h) and RyR-2 CRISPR/Cas9 Knockout CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RYR2 locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by RyR-2 CRISPR/Cas9 Knockout HDR Plasmid (h) and RyR-2 CRISPR/Cas9 Knockout HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RYR2 homology arms to support homology-directed repair at defined RYR2 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.