p38 alpha MAPK14 CRISPR Activation Plasmid (h): sc-400057-ACT

MAPK14 encodes p38 alpha, a stress-activated mitogen-activated protein kinase that integrates signals from cytokines, osmotic stress, UV irradiation, and pathogen-associated cues to regulate transcription, mRNA stability, and protein translation. p38 alpha MAPK14 functions within the MAPK cascade downstream of MKK3/MKK6 and modulates effectors such as ATF2, ELK1, MAPKAPK2/3, and HSP27, shaping inflammatory signaling, cell-cycle control, differentiation, and apoptosis. It is a key node in innate immune and inflammatory pathways, including TNF, IL-1, and TLR signaling, and influences crosstalk with NF-κB and interferon-responsive programs. Dysregulated MAPK14 activity has been implicated in chronic inflammation, neurodegenerative stress responses, cardiovascular remodeling, and multiple cancer-associated phenotypes, making it a widely studied target for pathway mapping and functional genomics.

p38 alpha MAPK14 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAPK14 expression without altering the underlying DNA sequence.

p38 alpha MAPK14 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAPK14 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 MAPK14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p38 alpha MAPK14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAPK14 locus and enabling the study of p38 alpha MAPK14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p38 alpha MAPK14 pathway restoration in tumor cells with silenced or reduced MAPK14 expression.

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