A-Myb CRISPR Activation Plasmid (h): sc-404328-ACT

MYBL1 encodes the human A-Myb transcription factor, a member of the MYB family that binds specific DNA motifs to regulate gene expression programs controlling cell cycle progression, differentiation, and lineage-specific proliferation. A-Myb activity intersects with transcriptional networks governing chromatin remodeling, replication-associated gene expression, and developmental signaling pathways, particularly in highly proliferative tissues. Dysregulated MYBL1 expression or rearrangement has been reported in multiple malignancies and is frequently studied for its impact on oncogenic transcription, tumor cell identity, and proliferative capacity. As an upstream regulator of gene networks, A-Myb provides a useful entry point for mechanistic studies of transcription factor dependency and pathway rewiring.

A-Myb CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYBL1 expression without altering the underlying DNA sequence.

A-Myb CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYBL1 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 MYBL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous A-Myb expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYBL1 locus and enabling the study of A-Myb-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of A-Myb pathway restoration in tumor cells with silenced or reduced MYBL1 expression.

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