APOBEC3G CRISPR Activation Plasmid (h): sc-402769-ACT

Human APOBEC3G is a cytidine deaminase that restricts retroviruses and retrotransposons by introducing C-to-U edits in single-stranded DNA during reverse transcription, promoting lethal mutagenesis of viral genomes. It functions within intrinsic innate immunity and intersects with interferon-stimulated gene programs and nucleic acid sensing pathways that shape antiviral defense. APOBEC3G activity is counteracted by viral antagonists such as HIV-1 Vif, making its regulation central to studies of host–pathogen interactions and viral evolution. Dysregulated APOBEC-family deaminase activity has also been linked to genome instability and mutation signatures relevant to cancer biology and inflammatory contexts.

APOBEC3G CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous APOBEC3G expression without altering the underlying DNA sequence.

APOBEC3G CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the APOBEC3G 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 APOBEC3G transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous APOBEC3G expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native APOBEC3G locus and enabling the study of APOBEC3G-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of APOBEC3G pathway restoration in tumor cells with silenced or reduced APOBEC3G expression.

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