RB1CC1 CRISPR Activation Plasmid (h): sc-416259-ACT

Human RB1CC1 (also known as FIP200) is a core component of the ULK1 autophagy initiation complex and coordinates early autophagosome formation in response to nutrient and energy cues. Through interactions with ULK1/2, ATG13, and ATG101, RB1CC1 links AMPK- and mTOR-regulated signaling to autophagic flux, while also contributing to cytoskeletal organization and selective cargo handling. RB1CC1-dependent autophagy influences mitochondrial quality control, proteostasis, and cellular stress adaptation. Dysregulation of RB1CC1-associated pathways has been implicated in contexts relevant to tumor biology and neurobiology, supporting its use as a mechanistic node for studying growth control, stress responses, and pathway crosstalk.

RB1CC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RB1CC1 expression without altering the underlying DNA sequence.

RB1CC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RB1CC1 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 RB1CC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RB1CC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RB1CC1 locus and enabling the study of RB1CC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RB1CC1 pathway restoration in tumor cells with silenced or reduced RB1CC1 expression.

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