LOC729974 CRISPR Activation Plasmid (h): sc-418913-ACT

RFPL4AL1 encodes the human protein LOC729974, a member of the RFPL family that is predicted to participate in nuclear protein–protein interactions and transcriptional regulation through zinc-finger/RING-like domains. Although the locus is not as extensively curated as canonical RFPL paralogs, RFPL-family genes have been associated with control of cell cycle progression, chromatin-associated processes, and context-dependent regulation of gene expression programs. Expression patterns reported for RFPL family members suggest roles in reproductive tissues and developmental signaling, providing a rationale for studying LOC729974 in lineage specification and differentiation models. Dysregulation of related RFPL genes has been observed in proliferative and oncogenic contexts, making LOC729974 a useful target for investigating transcriptional networks relevant to genome stability and aberrant growth phenotypes.

LOC729974 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RFPL4AL1 expression without altering the underlying DNA sequence.

LOC729974 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RFPL4AL1 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 RFPL4AL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LOC729974 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RFPL4AL1 locus and enabling the study of LOC729974-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LOC729974 pathway restoration in tumor cells with silenced or reduced RFPL4AL1 expression.

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