COLEC11 CRISPR Activation Plasmid (h): sc-413098-ACT

COLEC11 encodes collectin-11, a secreted C-type lectin pattern-recognition molecule that binds carbohydrate motifs on microbes and altered self-structures to initiate innate immune responses. Through interactions with MASP proteases, COLEC11 participates in lectin pathway complement activation, promoting opsonization and downstream inflammatory signaling. Beyond host defense, COLEC11 contributes to developmental processes such as cell migration and tissue patterning, consistent with its expression in multiple embryonic contexts. Dysregulated lectin pathway activity and COLEC11 variation have been associated with susceptibility to infectious and inflammatory conditions and with congenital developmental phenotypes, making it relevant for mechanistic studies of complement-mediated biology.

COLEC11 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COLEC11 expression without altering the underlying DNA sequence.

COLEC11 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COLEC11 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 COLEC11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COLEC11 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COLEC11 locus and enabling the study of COLEC11-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COLEC11 pathway restoration in tumor cells with silenced or reduced COLEC11 expression.

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