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A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells
Published on: December 12, 2017
Sindbis virus self-amplifying replicon is compatible with modified nucleotides mediating expression and vaccine
Hiva Azizi1, Gerard Agbayani1, Tyler M Renner1
1National Research Council Canada, Human Health Therapeutics, Ottawa, ON K1A 0R6, Canada.
Abstract:
The advent of effective formulations based on RNA encapsulated within lipid nanoparticles has led to the evaluation of this technology for its ability to treat and prevent a wide range of diseases. Expansion of the RNA-based tool kit available to vaccine and drug developers will accelerate this development and hopefully produce more robust and cost-effective therapies. With its more sustained expression profile, self-amplifying RNA offers many advantages to standard messenger RNA, allowing for the administration of lower RNA doses in vivo and potentially reducing dosing frequency for indications where long-term expression of a protein is required. We generated self-amplifying RNA containing the viral replicon from Sindbis virus, which demonstrated robust protein expression in vitro and in vivo. In addition, it showed increased compatibility with a number of modified nucleotides when compared to an equivalent RNA containing the replicon from Venezuelan equine encephalitis virus. These Sindbis-based self-amplifying RNAs containing modified nucleotides have the potential to be used in a variety of experimental and therapeutic applications.
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