LIF CRISPR Activation Plasmid (m): sc-421432-ACT

Mouse leukemia inhibitory factor (LIF) is a pleiotropic IL-6 family cytokine that signals through the LIF receptor and gp130 to activate JAK/STAT3, MAPK/ERK, and PI3K/AKT pathways. LIF regulates embryonic stem cell self-renewal and pluripotency, influences astrocyte differentiation and neuroinflammatory programs, and modulates hematopoietic and immune cell behavior. In peripheral tissues, LIF contributes to implantation and uterine receptivity, bone remodeling, and tissue stress responses by coordinating cytokine-driven transcriptional networks. Dysregulated LIF signaling has been linked to altered stem cell states, fibrotic remodeling, and tumor–stroma interactions in models of inflammation and cancer biology, supporting its use as a mechanistic node in pathway and phenotype studies.

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

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

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