LMX1B CRISPR Activation Plasmid (h): sc-402945-ACT

LMX1B encodes a LIM homeobox transcription factor that regulates lineage specification and organ morphogenesis, with prominent roles in dorsal limb patterning, renal glomerular development, and neuronal differentiation. In the nucleus, LMX1B binds DNA via its homeodomain and coordinates transcriptional programs through LIM domain–mediated protein interactions, shaping cell identity and developmental gene networks. Dysregulated LMX1B activity is linked to congenital syndromes involving nail, skeletal, and kidney abnormalities, and altered expression has been investigated in contexts of podocyte dysfunction and neurodevelopmental phenotypes. As a developmental regulator, LMX1B is widely used to study transcriptional control, enhancer logic, and cell fate transitions in relevant human cell models.

LMX1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LMX1B expression without altering the underlying DNA sequence.

LMX1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LMX1B 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 LMX1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LMX1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LMX1B locus and enabling the study of LMX1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LMX1B pathway restoration in tumor cells with silenced or reduced LMX1B expression.

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