RIN1 CRISPR Activation Plasmid (h): sc-402551-ACT

RIN1 (Ras and Rab interactor 1) is a Ras effector and Rab5 guanine nucleotide exchange factor that coordinates signal transduction with endocytic trafficking. By linking activated RAS to Rab5-dependent early endosome dynamics, RIN1 influences receptor internalization, downstream MAPK signaling amplitude, and cytoskeletal remodeling processes that shape cell migration and adhesion. RIN1 also interfaces with ABL family kinases and adaptor networks, positioning it within pathways that regulate membrane trafficking and actin-dependent responses. Dysregulated RIN1 expression or signaling context has been associated with altered growth factor receptor turnover and oncogenic signaling behaviors, making it relevant for mechanistic studies in cancer biology and cell signaling.

RIN1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RIN1 expression without altering the underlying DNA sequence.

RIN1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RIN1 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 RIN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RIN1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RIN1 locus and enabling the study of RIN1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RIN1 pathway restoration in tumor cells with silenced or reduced RIN1 expression.

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