Work

Efficient expression of a protein coding gene under the control of an RNA polymerase I promoter

Year: 1993

Type: article

Abstract: In mammalian cells, RNA polymerase I transcripts are uncapped and retain a polyphosphate 5' terminus. It is probably for this reason that they are poorly translated as messenger RNA. We show in this report that insertion of an Internal Ribosome Entry Site (IRES) into the 5' leader of an RNA polymerase I transcript overcomes the block to translation, presumably by substituting for the 5' trimethyl G cap. Addition of an SV40 polyA addition signal also enhances protein production from the RNA polymerase I transcript. RNA Polymerase I driven expression vectors containing both elements produce protein at levels comparable to that produced from RNA polymerase II driven expression vectors which utilize a retroviral LTR. RNA Polymerase I driven expression vectors may have a variety of uses both for basic research and for practical expression of recombinant proteins.

Source: Nucleic Acids Research

DOI: 10.1093/nar/21.15.3451

Authors: Theo D. Palmer, A.Dusty Miller, Ronald H. Reeder, Brian McStay

Institution: Fred Hutch Cancer Center

Language: English

Topic: RNA and protein synthesis mechanisms

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Institutes of Health, National Heart, Lung, and Blood Institute, National Institute of General Medical Sciences

Awards: CONTROL OF RIBOSOMAL GENE TRANSCRIPTION, HEMOPHILIA TREATMENT BY CLOTTING FACTOR GENE TRANSFER