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A self-initiating eukaryotic transient gene expression system based on contransfection of bacteriophage T7 RNA polymerase and DNA vectors containing a T7 autogene.
Nucleic Acids Res. 1994 Jun 11; 22(11):2114-20.NA

Abstract

A novel cytoplasmic gene expression system has been developed. This system differs from other expression systems in that it relies on the co-delivery of plasmid DNA and T7 RNA polymerase (RNAP) during transfection. The plasmid contains a T7 RNAP gene driven by the T7 promoter (T7 autogene) and a functional/reporter gene driven by another T7 promoter (T7T7/T7-gene construct). Once this DNA-enzyme complex is introduced into eukaryotic cells, the transcription of the T7 RNAP and the functional/reporter genes is initiated by the co-delivered T7 RNAP. The T7 RNAP, which is responsible for the initiation and maintenance of expression of both T7 and functional/reporter genes, is replenished by translation of newly synthesized T7 mRNA. This T7 system was designed in such a manner that the expression of the functional/reporter genes can occur in the cytoplasm and does not require any nuclear involvement. When transfected by either a pT7T7/T7Luc or a pT7T7/T7hGH plasmids with the cointroduced T7 RNAP, mouse L cells were found to express high levels of luciferase immediately after transfection, apparently due to the cytoplasmic gene expression; the expression of human growth hormone (hGH) could be sustained for at least 6 days. Both T7 and hGH mRNA were expressed by the cells transfected with pT7T7/T7hGH. These results suggest that this cytoplasmic expression system may be used for certain targets of somatic gene therapy.

Authors+Show Affiliations

Edison Biotechnology Institute, Ohio University, Athens 45701.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8029020

Citation

Chen, X, et al. "A Self-initiating Eukaryotic Transient Gene Expression System Based On Contransfection of Bacteriophage T7 RNA Polymerase and DNA Vectors Containing a T7 Autogene." Nucleic Acids Research, vol. 22, no. 11, 1994, pp. 2114-20.
Chen X, Li Y, Xiong K, et al. A self-initiating eukaryotic transient gene expression system based on contransfection of bacteriophage T7 RNA polymerase and DNA vectors containing a T7 autogene. Nucleic Acids Res. 1994;22(11):2114-20.
Chen, X., Li, Y., Xiong, K., & Wagner, T. E. (1994). A self-initiating eukaryotic transient gene expression system based on contransfection of bacteriophage T7 RNA polymerase and DNA vectors containing a T7 autogene. Nucleic Acids Research, 22(11), 2114-20.
Chen X, et al. A Self-initiating Eukaryotic Transient Gene Expression System Based On Contransfection of Bacteriophage T7 RNA Polymerase and DNA Vectors Containing a T7 Autogene. Nucleic Acids Res. 1994 Jun 11;22(11):2114-20. PubMed PMID: 8029020.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A self-initiating eukaryotic transient gene expression system based on contransfection of bacteriophage T7 RNA polymerase and DNA vectors containing a T7 autogene. AU - Chen,X, AU - Li,Y, AU - Xiong,K, AU - Wagner,T E, PY - 1994/6/11/pubmed PY - 1994/6/11/medline PY - 1994/6/11/entrez SP - 2114 EP - 20 JF - Nucleic acids research JO - Nucleic Acids Res. VL - 22 IS - 11 N2 - A novel cytoplasmic gene expression system has been developed. This system differs from other expression systems in that it relies on the co-delivery of plasmid DNA and T7 RNA polymerase (RNAP) during transfection. The plasmid contains a T7 RNAP gene driven by the T7 promoter (T7 autogene) and a functional/reporter gene driven by another T7 promoter (T7T7/T7-gene construct). Once this DNA-enzyme complex is introduced into eukaryotic cells, the transcription of the T7 RNAP and the functional/reporter genes is initiated by the co-delivered T7 RNAP. The T7 RNAP, which is responsible for the initiation and maintenance of expression of both T7 and functional/reporter genes, is replenished by translation of newly synthesized T7 mRNA. This T7 system was designed in such a manner that the expression of the functional/reporter genes can occur in the cytoplasm and does not require any nuclear involvement. When transfected by either a pT7T7/T7Luc or a pT7T7/T7hGH plasmids with the cointroduced T7 RNAP, mouse L cells were found to express high levels of luciferase immediately after transfection, apparently due to the cytoplasmic gene expression; the expression of human growth hormone (hGH) could be sustained for at least 6 days. Both T7 and hGH mRNA were expressed by the cells transfected with pT7T7/T7hGH. These results suggest that this cytoplasmic expression system may be used for certain targets of somatic gene therapy. SN - 0305-1048 UR - https://www.unboundmedicine.com/medline/citation/8029020/A_self_initiating_eukaryotic_transient_gene_expression_system_based_on_contransfection_of_bacteriophage_T7_RNA_polymerase_and_DNA_vectors_containing_a_T7_autogene_ L2 - https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/22.11.2114 DB - PRIME DP - Unbound Medicine ER -