Mcl-1 CRISPR Activation Plasmid (m): sc-421589-ACT

Mouse Mcl1 encodes Mcl-1, an anti-apoptotic BCL-2 family protein that preserves mitochondrial outer membrane integrity by sequestering pro-death BH3-only factors and restraining BAX/BAK activation. Its abundance is tightly controlled by growth factor signaling, stress responses, and proteasomal turnover, positioning Mcl-1 as a rapid-response node in intrinsic apoptosis. Mcl-1 also interfaces with cellular programs including mitochondrial metabolism, cell-cycle progression, and survival downstream of pathways such as PI3K–AKT and MAPK/ERK. Dysregulated Mcl1 expression is frequently associated with altered apoptotic thresholds that contribute to aberrant cell survival in cancer biology and to survival phenotypes in immune and hematopoietic lineages, making it a key target for mechanistic studies of stress adaptation and cell fate.

Mcl-1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Mcl1 expression without altering the underlying DNA sequence.

Mcl-1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Mcl1 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 Mcl1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Mcl-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Mcl1 locus and enabling the study of Mcl-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Mcl-1 pathway restoration in tumor cells with silenced or reduced Mcl1 expression.

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