EKLF/KLF1 CRISPR Activation Plasmid (h): sc-402366-ACT

KLF1 (erythroid Krüppel-like factor; EKLF) is a lineage-restricted transcription factor that coordinates erythropoiesis by controlling stage-specific gene expression programs in erythroid progenitors. It binds CACCC elements and cooperates with core erythroid regulators to promote hemoglobin switching, membrane and cytoskeletal maturation, and erythroid metabolic remodeling, integrating with GATA1/TAL1-driven transcriptional networks. KLF1 also influences chromatin state through recruitment of co-regulators, shaping enhancer activity and erythroid differentiation trajectories. Dysregulation or mutation of KLF1 is linked to inherited red blood cell phenotypes, including altered globin expression and anemia-associated traits, making it a key node for studying erythroid gene regulation and hematologic disease mechanisms.

EKLF/KLF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KLF1 expression without altering the underlying DNA sequence.

EKLF/KLF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KLF1 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 KLF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EKLF/KLF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KLF1 locus and enabling the study of EKLF/KLF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EKLF/KLF1 pathway restoration in tumor cells with silenced or reduced KLF1 expression.

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