FAM150B CRISPR Activation Plasmid (h): sc-409087-ACT

FAM150B (also known as ALKAL2) encodes a secreted ligand implicated in activation of the ALK family of receptor tyrosine kinases, supporting cell–cell communication and context-dependent regulation of growth and differentiation programs. Through ALK signaling, FAM150B can intersect with canonical downstream cascades such as MAPK/ERK and PI3K/AKT that influence proliferation, survival, and developmental patterning. Expression of FAM150B is observed in neural and other tissues, and dysregulated ALK-axis signaling is relevant to oncogenic and neurodevelopmental research contexts. As a modulator of receptor tyrosine kinase activity, FAM150B provides a useful node for studying extracellular control of signaling thresholds, receptor activation dynamics, and transcriptional responses.

FAM150B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FAM150B expression without altering the underlying DNA sequence.

FAM150B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FAM150B 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 FAM150B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FAM150B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FAM150B locus and enabling the study of FAM150B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FAM150B pathway restoration in tumor cells with silenced or reduced FAM150B expression.

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