LIN-9 CRISPR Activation Plasmid (h): sc-418584-ACT

Human LIN9 encodes LIN-9, a core component of the MuvB/DREAM regulatory network that coordinates cell-cycle gene expression by switching between transcriptional repression in quiescence and activation during S phase and mitosis. LIN-9 helps assemble complexes with E2F and B-Myb/FOXM1-associated factors to regulate promoters of genes controlling DNA replication, mitotic spindle dynamics, and cytokinesis. Through these interactions, LIN-9 contributes to genome stability and orderly cell-cycle progression, making it relevant to studies of proliferative dysregulation and chromosomal instability in cancer biology. LIN9-dependent transcriptional programs are also used to interrogate checkpoints and stress responses that influence proliferation in diverse cell types.

LIN-9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LIN9 expression without altering the underlying DNA sequence.

LIN-9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LIN9 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 LIN9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LIN-9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LIN9 locus and enabling the study of LIN-9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LIN-9 pathway restoration in tumor cells with silenced or reduced LIN9 expression.

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