Su[fu] CRISPR/Cas9 KO Plasmid (m): sc-423973

Overview

Suppressor of fused (Sufu) encodes the Su(fu) protein, a key negative regulator of Hedgehog signaling that restrains GLI transcription factor activity and helps control transcriptional programs governing embryonic patterning, cell fate specification, and tissue homeostasis. In mouse cells, SUFU functions as a cytoplasmic scaffold that sequesters GLI proteins and coordinates their processing and nuclear access, integrating with primary cilium–dependent Hedgehog pathway dynamics. Perturbation of Sufu can shift pathway output and downstream gene expression, making it relevant for studying developmental signaling logic and Hedgehog-driven cellular phenotypes. Altered SUFU function has been associated with dysregulated Hedgehog signaling in disease-related models, supporting its use as a mechanistic node for pathway interrogation.

Su[fu] CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Sufu gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Sufu 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 Sufu 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 Su[fu] protein expression.

This CRISPR knockout system enables efficient generation of Sufu-deficient cell models for investigation of Su[fu] signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting Sufu exon(s) critical for Su[fu] 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 Sufu genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by Su[fu] CRISPR/Cas9 KO Plasmid (m) and Su[fu] CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Sufu locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by Su[fu] HDR Plasmid (m) and Su[fu] HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Sufu homology arms to support homology-directed repair at defined Sufu 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.