Lamin B2 CRISPR Activation Plasmid (h): sc-401873-ACT

LMNB2 encodes lamin B2, a core component of the nuclear lamina that polymerizes beneath the inner nuclear membrane to maintain nuclear shape and mechanical stability. Lamin B2 participates in chromatin tethering, DNA replication timing, and nuclear envelope reassembly during mitosis, linking genome organization to cell cycle progression and stress responses. Through interactions with lamin-associated domains and nuclear pore complexes, it influences transcriptional programs and genome integrity pathways. Altered lamin B family function is associated with nuclear architecture defects, aberrant chromatin organization, and phenotypes relevant to premature aging, neurodevelopment, and cancer cell biology.

Lamin B2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LMNB2 expression without altering the underlying DNA sequence.

Lamin B2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LMNB2 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 LMNB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Lamin B2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LMNB2 locus and enabling the study of Lamin B2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Lamin B2 pathway restoration in tumor cells with silenced or reduced LMNB2 expression.

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