title: Next generation sequencing of DNA-launched Chikungunya vaccine virus.
abstract: Chikungunya virus (CHIKV) represents a pandemic threat with no approved vaccine available. Recently, we described a novel vaccination strategy based on iDNA® infectious clone designed to launch a live-attenuated CHIKV vaccine from plasmid DNA in vitro or in vivo. As a proof of concept, we prepared iDNA plasmid pCHIKV-7 encoding the full-length cDNA of the 181/25 vaccine. The DNA-launched CHIKV-7 virus was prepared and compared to the 181/25 virus. Illumina HiSeq2000 sequencing revealed that with the exception of the 3' untranslated region, CHIKV-7 viral RNA consistently showed a lower frequency of single-nucleotide polymorphisms than the 181/25 RNA including at the E2-12 and E2-82 residues previously identified as attenuating mutations. In the CHIKV-7, frequencies of reversions at E2-12 and E2-82 were 0.064% and 0.086%, while in the 181/25, frequencies were 0.179% and 0.133%, respectively. We conclude that the DNA-launched virus has a reduced probability of reversion mutations, thereby enhancing vaccine safety.
authors: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD 21701, USA.
authors: Rachmat
authors: R
authors: Hidajat
authors: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD 21701, USA.
authors: Brian
authors: B
authors: Nickols
authors: Institute for Human Infections and Immunity, Sealy Center for Vaccine Development and Department of Pathology, University of Texas Medical Branch, GNL, 301 University Blvd., Galveston, TX 77555, USA.
authors: Naomi
authors: N
authors: Forrester
authors: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD 21701, USA.
authors: Irina
authors: I
authors: Tretyakova
authors: Institute for Human Infections and Immunity, Sealy Center for Vaccine Development and Department of Pathology, University of Texas Medical Branch, GNL, 301 University Blvd., Galveston, TX 77555, USA.
authors: Scott
authors: S
authors: Weaver
authors: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD 21701, USA. Electronic address: ppushko@medigen-usa.com.
authors: Peter
authors: P
authors: Pushko
mesh_terms: D001483:Base Sequence; D065632:Chikungunya Fever / Q000821:virology*; D002646:Chikungunya virus / Q000235:genetics* / Q000302:isolation & purification; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines / Q000235:genetics*
chemical_list: D019444:Vaccines, DNA; D014765:Viral Vaccines
keywords: Alphavirus; CHIKV; Chikungunya fever; Chikungunya virus; DNA vaccine; Live attenuated vaccine