βB1-crystallin CRISPR Activation Plasmid (h): sc-403361-ACT

CRYBB1 encodes βB1-crystallin, a major structural component of the human eye lens that contributes to high protein packing density, refractive index, and long-term transparency. βB1-crystallin participates in crystallin complex assembly and lens fiber cell maturation, where proteostasis and resistance to aggregation are essential for maintaining optical clarity. Dysregulation of CRYBB1 expression or protein stability has been associated with inherited lens opacities and cataract-related phenotypes, making it a useful target for studying protein aggregation, lens development, and stress-response pathways in ocular models. Research on CRYBB1 also informs mechanisms of long-lived protein maintenance, post-translational modification, and chaperone interactions in terminally differentiated tissues.

βB1-crystallin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CRYBB1 expression without altering the underlying DNA sequence.

βB1-crystallin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CRYBB1 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 CRYBB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous βB1-crystallin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CRYBB1 locus and enabling the study of βB1-crystallin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of βB1-crystallin pathway restoration in tumor cells with silenced or reduced CRYBB1 expression.

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