NEIL2 CRISPR Activation Plasmid (h): sc-408091-ACT

NEIL2 (endonuclease VIII-like 2) is a DNA glycosylase/AP lyase that initiates base excision repair by recognizing and excising oxidized DNA lesions, including those arising in single-stranded, transcription-associated, and replicating DNA. By coupling lesion recognition to strand incision, NEIL2 helps maintain genome stability and modulates transcriptional fidelity under oxidative stress, intersecting functionally with BER factors such as APEX1, POLB, XRCC1, and downstream ligation steps. NEIL2 activity is linked to cellular responses to reactive oxygen species and inflammation, processes that shape mutational burden and stress signaling in many biological contexts. Dysregulated oxidative DNA repair capacity involving NEIL2 has been studied in relation to cancer-associated genomic instability and chronic inflammatory disease mechanisms, supporting its relevance in DNA damage biology.

NEIL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NEIL2 expression without altering the underlying DNA sequence.

NEIL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NEIL2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the NEIL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NEIL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NEIL2 locus and enabling the study of NEIL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NEIL2 pathway restoration in tumor cells with silenced or reduced NEIL2 expression.

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