TCL-1A CRISPR Activation Plasmid (h): sc-402028-ACT

TCL1A encodes the human T-cell leukemia/lymphoma 1A protein (TCL-1A), a cytoplasmic regulator of kinase signaling that can potentiate PI3K–AKT pathway activity through protein–protein interactions. TCL-1A is most prominently expressed in early lymphoid compartments and contributes to lymphocyte development, survival, and metabolic signaling programs. Dysregulated TCL1A expression has been linked to aberrant B- and T-cell proliferation and is frequently studied in the context of hematologic malignancies, where altered AKT-dependent transcriptional and apoptotic control can impact oncogenic phenotypes. As a pathway-relevant modulator, TCL1A is commonly interrogated to map upstream regulators and downstream effectors of AKT signaling in immune cells.

TCL-1A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TCL1A expression without altering the underlying DNA sequence.

TCL-1A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TCL1A 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 TCL1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TCL-1A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TCL1A locus and enabling the study of TCL-1A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TCL-1A pathway restoration in tumor cells with silenced or reduced TCL1A expression.

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