CAML CRISPR Activation Plasmid (h): sc-403690-ACT

CAMLG encodes calcium-modulating cyclophilin ligand (CAML), an integral membrane protein of the endoplasmic reticulum that couples intracellular Ca²⁺ signaling to membrane protein biogenesis and cell survival programs. CAML participates in receptor- and store-operated calcium entry processes and has been linked to regulation of lymphocyte activation, apoptosis control, and proteostasis through ER-associated pathways. By influencing calcium-dependent transcriptional responses and stress signaling, CAML can modulate proliferation and immune cell function. Dysregulation of CAMLG expression or CAML-mediated calcium handling has been associated with altered survival signaling and oncogenic phenotypes in experimental models, supporting its relevance for mechanistic studies in cancer and immune biology.

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

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

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