論文

国際誌
2021年4月20日

Establishment of a reverse genetics system for SARS-CoV-2 using circular polymerase extension reaction.

Cell reports
  • Shiho Torii
  • ,
  • Chikako Ono
  • ,
  • Rigel Suzuki
  • ,
  • Yuhei Morioka
  • ,
  • Itsuki Anzai
  • ,
  • Yuzy Fauzyah
  • ,
  • Yusuke Maeda
  • ,
  • Wataru Kamitani
  • ,
  • Takasuke Fukuhara
  • ,
  • Yoshiharu Matsuura

35
3
開始ページ
109014
終了ページ
109014
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.celrep.2021.109014

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the causative agent of coronavirus disease 2019 (COVID-19). Although multiple mutations have been observed in SARS-CoV-2, functional analysis of each mutation of SARS-CoV-2 has been limited by the lack of convenient mutagenesis methods. In this study, we establish a PCR-based, bacterium-free method to generate SARS-CoV-2 infectious clones. Recombinant SARS-CoV-2 could be rescued at high titer with high accuracy after assembling 10 SARS-CoV-2 cDNA fragments by circular polymerase extension reaction (CPER) and transfection of the resulting circular genome into susceptible cells. The construction of infectious clones for reporter viruses and mutant viruses could be completed in two simple steps: introduction of reporter genes or mutations into the desirable DNA fragments (∼5,000 base pairs) by PCR and assembly of the DNA fragments by CPER. This reverse genetics system may potentially advance further understanding of SARS-CoV-2.

リンク情報
DOI
https://doi.org/10.1016/j.celrep.2021.109014
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/33838744
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8015404
ID情報
  • DOI : 10.1016/j.celrep.2021.109014
  • PubMed ID : 33838744
  • PubMed Central 記事ID : PMC8015404

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