C9orf152 CRISPR/Cas9 KO Plasmid (h): sc-415979

Overview

C9orf152 encodes a poorly characterized human open reading frame protein with limited functional annotation and emerging evidence for context-dependent expression across tissues and cell states. Available transcriptomic and proteomic datasets suggest potential involvement in fundamental cellular programs such as transcriptional regulation, protein homeostasis, or subcellular organization, although direct pathway placement remains to be fully defined. As with many minimally annotated loci, C9orf152 is of interest for systems-level studies linking gene expression signatures to cellular phenotypes and for interpreting association signals in genomic datasets. Ongoing functional genomics efforts can help clarify whether C9orf152 contributes to disease-relevant processes through effects on proliferation, stress responses, or differentiation.

C9orf152 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the C9orf152 gene in human cell lines. Each plasmid in the pool co-expresses a unique sgRNA, targeting a distinct site within the C9orf152 locus, alongside the Streptococcus pyogenes Cas9 nuclease, and encodes GFP to enable fluorescent identification and enrichment of successfully transfected cells. This multi-guide strategy increases the likelihood of inducing frameshifts or deletions that produce a functional knockout, offering a more robust alternative to single-guide approaches. DSBs induced at multiple sites are resolved through non-homologous end joining (NHEJ) or, when used with the included HDR donor template, homology-directed repair (HDR) at a defined target site within the locus.

When used in conjunction with the RFP-expressing HDR donor, GFP and RFP fluorescence can be used together to distinguish transfected from edited cell populations, streamlining flow cytometry-based sorting and clone selection workflows.

Homology-Directed Repair (HDR) Donor — Puromycin Cassette with RFP Reporter

For applications requiring confirmed, selectable knockout clones, C9orf152 HDR Plasmid (h) includes an HDR donor construct containing a puromycin resistance cassette (PuroR) and a red fluorescent protein (RFP) reporter, flanked by homology arms specific to a defined C9orf152 target site.
When co-transfected with C9orf152 CRISPR/Cas9 KO Plasmid (h):

• The PuroR-RFP cassette integrates at the Cas9 cut site via HDR, disrupting the C9orf152 open reading frame.
• RFP fluorescence provides an immediate visual indicator of successful integration, enabling fluorescence-based identification or sorting of edited cells prior to or alongside puromycin selection.
• Successfully edited cells are confirmed through puromycin resistance, substantially reducing clone screening burden.
• This selection strategy is ideal for generating stable, clonal KO cell lines for downstream functional studies, drug screening, or model development.

Cre-lox Cassette Removal System

The HDR donor construct features loxP sites flanking the PuroR-RFP selection cassette to allow clean marker removal following clone confirmation. Transient expression of Cre recombinase via the included Cre Vector: sc-418923 excises the cassette, leaving a minimal residual loxP site within the C9orf152 locus and eliminating potential confounding effects on downstream assays.
This two-step approach:

• Minimizes disruption to local chromatin architecture and neighboring regulatory elements
• Restores a near-native genomic context at the edited locus
• Enables reuse of the puromycin selection strategy in the same cell line for additional edits

Key Features

• gRNA targeting C9orf152 exon(s) critical for C9orf152 function
• Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
• HDR donor with puromycin resistance for positive clone selection
• loxP-flanked PuroR cassette with Cre recombinase vector for seamless marker removal
• Supplied ready to use for delivery by transfection

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.