Riboflavin kinase CRISPR/Cas9 KO Plasmid (h): sc-418359

Overview

Human RFK encodes riboflavin kinase, a cytosolic enzyme that phosphorylates riboflavin (vitamin B2) to flavin mononucleotide (FMN), supplying a key precursor for flavin adenine dinucleotide (FAD) biosynthesis. By controlling intracellular FMN/FAD availability, RFK supports flavoprotein-dependent redox reactions that impact mitochondrial oxidative metabolism, antioxidant defense, and broader cellular bioenergetics. Perturbation of riboflavin utilization and flavin cofactor homeostasis has been associated with metabolic dysfunction and oxidative stress–linked phenotypes, making RFK a useful node for studying vitamin-dependent enzymology. RFK function is therefore relevant to investigations of mitochondrial processes, redox signaling, and nutrient–cofactor regulation in human cell models.

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

This CRISPR knockout system enables efficient generation of RFK-deficient cell models for investigation of Riboflavin kinase signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting RFK exon(s) critical for Riboflavin kinase 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 RFK genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

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