TTF-1/Thyroid Transcription Factor 1/NKX2-1 CRISPR Activation Plasmid (h): sc-400458-ACT

NKX2-1 encodes the homeobox transcription factor TTF-1 (also known as Thyroid Transcription Factor 1), a lineage-determining regulator of endoderm-derived organs that coordinates transcriptional programs required for thyroid, lung, and forebrain development and differentiation. In the thyroid, NKX2-1 controls expression of genes involved in hormone biosynthesis and epithelial polarity, while in pulmonary epithelium it regulates surfactant homeostasis and alveolar maturation. NKX2-1 integrates with other developmental pathways, including WNT, BMP, and FGF signaling, to establish and maintain tissue-specific gene expression states. Dysregulated NKX2-1 activity is associated with congenital thyroid and respiratory phenotypes and is frequently used as a molecular marker and functional node in studies of thyroid and lung disease biology, including lineage plasticity and oncogenic transcriptional networks.

TTF-1/Thyroid Transcription Factor 1/NKX2-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NKX2-1 expression without altering the underlying DNA sequence.

TTF-1/Thyroid Transcription Factor 1/NKX2-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NKX2-1 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 NKX2-1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TTF-1/Thyroid Transcription Factor 1/NKX2-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NKX2-1 locus and enabling the study of TTF-1/Thyroid Transcription Factor 1/NKX2-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TTF-1/Thyroid Transcription Factor 1/NKX2-1 pathway restoration in tumor cells with silenced or reduced NKX2-1 expression.

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