Next generation sequencing of DNA-launched Chikungunya vaccine virus

作者：Hidajat Rachmat; Nickols Brian; Forrester Naomi; Tretyakova Irina; Weaver Scott; Pushko Peter^*

来源：Virology, 2016, 490: 83-90.

DOI：10.1016/j.virol.2016.01.009

• Chikungunya virus
• Chikungunya fever
• CHIKV
• DNA vaccine
• Live attenuated vaccine
• Alphavirus

Chikungunya virus (CHIKV) represents a pandemic threat with no approved vaccine available. Recently, we described a novel vaccination strategy based on iDNA (R) 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.

• 出版日期2016-3