MNDA CRISPR/Cas9 KO Plasmid (h): sc-406492

Overview

Myeloid cell nuclear differentiation antigen (MNDA) is an interferon-inducible nuclear protein predominantly expressed in granulocytes and monocytes that participates in innate immune regulation and myeloid differentiation programs. MNDA has been linked to control of inflammatory gene expression and antimicrobial responses, functioning within interferon-stimulated gene networks and broader transcriptional regulatory processes in the nucleus. Altered MNDA expression is frequently used as a marker of myeloid lineage state and has been associated with immune dysregulation in hematologic disease contexts, including leukemic phenotypes and aberrant inflammatory signaling. As part of the p200/IFI family, MNDA is studied for its contributions to host defense pathways, cytokine-driven activation states, and cell fate decisions in myeloid compartments.

MNDA CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the MNDA gene in human cell lines. Each plasmid in the pool co-expresses a unique sgRNA, targeting a distinct site within the MNDA 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, MNDA 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 MNDA target site.
When co-transfected with MNDA CRISPR/Cas9 KO Plasmid (h):

• The PuroR-RFP cassette integrates at the Cas9 cut site via HDR, disrupting the MNDA 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 MNDA 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 MNDA exon(s) critical for MNDA 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.