Abstract: Objective To construct rat N1ICD lentiviral over-expression vector (LV-N1ICD) and N1ICD lentivirus interference
vector (LV-N1ICD-shRNA. Methods With the rat cDNA as a template, the N1ICD fragment was amplified by PCR to construct
pGC-FU-N1ICD-3Flag shuttle plasmid by directly clone. Four pairs of N1ICD-shRNA oligonucleotide sequences were synthesized
to construct the GVC112-N1ICD-shRNA interference plasmid. pGC-FU-N1ICD-3Flag and GVC112 -N1ICD-shRNA
plasmids were co-transfected into 293T cells to screen for the best interference plasmid in the 4 GVC112-N1ICD-shRNA
plasmids by detecting Flag expression. pGC-FU-N1ICD-3Flag or GVC112-N1ICD-shRNA plasmid along with with pHelper 1.0
and pHelper 2.0 plasmids were co-transfect into 293T cells to package LV-N1ICD and LV-N1ICD-shRNA, and the virus titer
was determined by real-time PCR and drug screening method, respectively. H9c2 cells infected with LV-N1ICD and LVN1ICD-
shRNA respectively were assessed for cell viability using CCK-8 assay. Results pGC-FU-N1ICD-3Flag and GVC112-
N1ICD-shRNA plasmid were verified by PCR, gene sequencing and Western blotting. Co-transfection of the plasmids with
pHelper 1.0, and pHelper 2.0 plasmids into 293T cells obtained high-titer LV-N1ICD and LV-N1ICD-shRNA. LV-N1ICD was
capable of promoting the cell viability and LV-N1ICD-shRNA produced an opposite effect. Conclusion The vectors LV-N1ICD
and LV-N1ICD-shRNA have been successfully constructed and packaged, which have the biological functions of Notch1
signaling.