Mcl-1 CRISPR Activation Plasmid (h): sc-400079-ACT

MCL1 encodes Mcl-1, an anti-apoptotic BCL-2 family protein that localizes primarily to the outer mitochondrial membrane and suppresses mitochondrial outer membrane permeabilization by sequestering pro-apoptotic BH3-only factors and inhibiting BAX/BAK activation. Its abundance is tightly regulated through rapid turnover and signaling inputs from pathways such as PI3K–AKT, MAPK/ERK, and JAK/STAT, enabling dynamic control of intrinsic apoptosis during proliferation, differentiation, and cellular stress responses. Mcl-1 also contributes to mitochondrial homeostasis and has been linked to the regulation of autophagy and metabolic adaptation. Dysregulated MCL1 expression is frequently associated with altered cell survival programs in cancer and influences resistance to genotoxic and targeted perturbations, making it a key node for studying apoptosis, stress signaling, and survival dependencies.

Mcl-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MCL1 expression without altering the underlying DNA sequence.

Mcl-1 CRISPR Activation Plasmid (h) 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.