NOTCH1-ICD ORF cDNA Lentivirus

Background

NOTCH1 is a single-pass transmembrane receptor that mediates short-range, cell-to-cell signaling central to development and tissue homeostasis. Ligand binding triggers proteolytic cleavage of the receptor and release of the NOTCH1 intracellular domain (NOTCH1-ICD), which translocates to the nucleus and partners with the CSL/RBPJ transcription factor and Mastermind co-activators to activate target genes such as HES and HEY family members. NOTCH1 signaling governs cell-fate decisions, differentiation, and proliferation in many lineages. Aberrant NOTCH1 activity is implicated in human disease, most notably as an oncogenic driver in T-cell acute lymphoblastic leukemia, making the constitutively active intracellular domain a valuable research tool.

Product Description & Applications

This ORF cDNA lentivirus drives stable overexpression of the human NOTCH1 intracellular domain (NOTCH1-ICD), the constitutively active fragment of the receptor, in mammalian cells. Expression is driven by a CMV promoter, and individual configurations may include a C-terminal epitope tag, a fluorescent reporter such as EGFP or mCherry, and an antibiotic selection marker, with elements linked by self-cleaving peptide sequences for independent translation from a single transcript.

Delivered as a third-generation, VSV-G-pseudotyped system, the particles transduce a wide range of cells, including primary and thawed cultures, and support establishment of long-term stable lines by antibiotic selection or FACS. The product is used to study constitutive NOTCH1 pathway activation in cell-fate, differentiation, and cancer research.

About This Product

This ORF cDNA lentivirus enables stable overexpression of NOTCH1 (NCBI Accession: NM_0176170) in mammalian cells via a third-generation, VSV-G pseudotyped delivery system. The ORF cDNA is fused to a C-terminal epitope tag (V5, Myc, or HA) and expressed under a strong constitutive promoter (CMV). Reporter and selection marker components (EGFP, GFP, mCherry, RFP; Blasticidin, Puromycin) are co-expressed via self-cleaving P2A peptides, enabling independent protein production without fusion-tag artifacts.

Ultra-purification by PEG precipitation and sucrose gradient centrifugation yields high-titer particles suitable for primary cells, suspension cultures, and stem cells. Stable polyclonal cell lines are established within 10–14 days by antibiotic selection or FACS sorting. For in vivo applications, the serum-free formulation and VSV-G envelope support direct administration or further concentration for stereotactic injection.