RB1CC1 CRISPR Activation Plasmid (m): sc-419502-ACT

Rb1cc1 encodes RB1CC1 (also known as FIP200), a core component of the ULK1 autophagy-initiating complex that coordinates autophagosome formation in response to nutrient status and cellular stress. RB1CC1 also interfaces with mTOR and AMPK signaling to couple growth-factor and energy-sensing pathways to autophagic flux, influencing mitochondrial quality control and proteostasis. Through these roles, RB1CC1 impacts cell survival, inflammation, and tissue homeostasis, making it widely studied in contexts such as neurodegeneration, metabolic dysregulation, and cancer biology. In mouse models, modulation of RB1CC1 provides a mechanistic handle to interrogate stress-adaptation programs and autophagy-dependent remodeling in diverse cell types.

RB1CC1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Rb1cc1 expression without altering the underlying DNA sequence.

RB1CC1 CRISPR Activation Plasmid (m) 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.