A novel inducible lentiviral system for multi-gene expression with human HSP70 promoter and tetracycline-induced promoter

Abstract

Despite lentiviral system's predominance, its ultimate potential for gene therapy has not been fully exploited. Currently, most lentivirus vectors are non-inducible expression system or single-gene-induced system, which limits the extensive application in gene therapy. In this study, we designed a novel lentiviral vector containing HSP70 promoter and TRE promoter. Compared to traditional lentiviral vectors and inducible vectors, our controllable system has many advantages. Firstly, it contains multiple gene or shRNA targets. Secondly, genes expression is on/off in response to heat shock and DOX induction in time of need respectively with high effectivity and sensitivity. Thirdly, TRE promoter and HSP70 promoter can work with no interference from each other in the same inducible lentiviral vector. In addition, our study also shows that our novel vector has a higher downstream gene expression efficiency than co-transfection method and can co-position multi-genes in single cell effectively. Finally, we propose four derived models based on our vector at the end, which may be useful in biological research and clinical research in the future. Therefore, we believe that this novel lentiviral system could be promising in gene therapy for tumor.

MDM2 induces EMT via the B‑Raf signaling pathway through 14‑3‑3

“…Plasmid vector construction. Plasmid construction was performed as described in our previous studies (37)(38)(39). pLenti-CMV-MDM2-green fluorescent protein (GFP)-puro was constructed based on a plenti-CMV-puro backbone.…”

Optogenetic Control of Programmable Genome Editing by Photoactivatable CRISPR/Cas9 Nanosystem in the Second Near-Infrared Window

“…To construct HSP70-Cas9-GFP-luciferase-U6-sgRNA plasmid, HSP70 promoter core sequence 66 was amplified from genomic DNA of 293T cells by PCR and inserted into pCS2-CMV-Cas9-P2A-GFP-P2A-luciferase-pA (Addgene #48138) by replacing CMV promoter through Hind III restriction site. U6-sgRNA fragment was also added to the plasmid by SalI.…”

A logic “OR” gate composed of a blue light‐induced CRISPR/dCas9 system and a Tet‐on system activates cancer suppressor genes to inhibit the growth of cutaneous squamous cell carcinoma cells

“…[22] In this study, we applied the principle of synthetic biology and combined a light-induced gene circuit with a tetracycline-induced gene circuit to construct the logical "OR" gate of tumour killing, and studied the synergistic effect of the two gene circuits on tumour killing. On the basis of previous studies, [13][14][15][16][17][18][19][20][21][22][23][24] the possibility of tumour therapy in clinical application was discussed more deeply according to the characteristics of special types of tumours.…”

“…[15] An inducible gene expression system can regulate gene expression in time and space, which is one of the important tools for gene function research, among which the tetracycline-induced gene expression system is the most widely used. [16][17][18][19][20] The tetracycline-induced gene expression system consists of a regulatory expression vector and a reactive expression vector. [21] The regulatory expression vector contains a human cytomegalovirus early promoter (PhCMV) and antisense Tet transcriptional activator (reverse tetracycline tranional activator, rtTA).…”