RNA interference as a metabolic engineering tool: potential for in vivo control of protein expression in an insect larval model.
Many ex vivo factors influence the yield of recombinant protein produced via AcMNPV (Autographa californica multiple nucleocapsid nuclear polyhedrosis virus) in Trichoplusia ni (T. ni) larvae. Among these are: the method of infection, the time of infection, the virus load, and the time of harvest. In vivo strategies, however, that attempt to manipulate host function in this and other expression systems have largely been ignored. In this work, RNA interference (RNAi) is shown as an effective metabolic engineering controller to downregulate targeted gene expression. Specifically, RNAi was made to virus-encoded gfp(uv) and was found to inhibit the production of GFPuv in larvae when injected within an 18-h window (before and after) of baculovirus infection. The level of inhibition was found to depend, both in duration and extent, on the concentration of injected RNAi. That relatively low levels of RNAi can inhibit protein synthesis driven by the strong polyhedrin (polh) promoter of AcMNPV, suggests that RNAi will find utility as an in vivo metabolic controller in metabolic engineering studies such as this one pertaining to protein expression.
Kramer
Shannon F
SF
Center for Biosystems Research, University of Maryland Biotechnology Institute, 5115 Plant Sciences Building, College Park, MD 20742, USA.
Bentley
William E
WE
D000818:Animals; D005786:Gene Expression Regulation / Q000235:genetics; D049452:Green Fluorescent Proteins; D007814:Larva / Q000235:genetics / Q000378:metabolism; D007915:Lepidoptera / Q000235:genetics* / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D034622:RNA Interference / Q000502:physiology*; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism*; D014162:Transfection / Q000379:methods
D008164:Luminescent Proteins; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins