Controlling the Replication of a Genomically Recoded HIV-1 with a Functional Quadruplet Codon in Mammalian Cells

Yan Chen, Yanmin Wan, Nanxi Wang, Zhe Yuan, Wei Niu, Qingsheng Li, Jiantao Guo

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Large efforts have been devoted to genetic code engineering in the past decade, aiming for unnatural amino acid mutagenesis. Recently, an increasing number of studies were reported to employ quadruplet codons to encode unnatural amino acids. We and others have demonstrated that the quadruplet decoding efficiency could be significantly enhanced by an extensive engineering of tRNAs bearing an extra nucleotide in their anticodon loops. In this work, we report the identification of tRNA mutants derived from directed evolution to efficiently decode a UAGA quadruplet codon in mammalian cells. Intriguingly, the trend of quadruplet codon decoding efficiency among the tested tRNA variants in mammalian cells was largely the same as that in E. coli. We subsequently demonstrate the utility of quadruplet codon decoding by the construction of the first HIV-1 mutant that lacks any in-frame amber nonsense codons and can be precisely activated by the decoding of a genomically embedded UAGA codon with an unnatural amino acid. Such conditionally activatable HIV-1 mutant can likely facilitate both fundamental investigations of HIV-1 as well as vaccine developments. The use of quadruplet codon, instead of an amber nonsense codon, to control HIV-1 replication has the advantage in that the correction of a frameshift caused by a quadruplet codon is much less likely than the reversion of an amber codon back into a sense codon in HIV-1.

Original languageEnglish (US)
Pages (from-to)1612-1617
Number of pages6
JournalACS Synthetic Biology
Volume7
Issue number6
DOIs
StatePublished - Jun 15 2018

Fingerprint

Codon
Decoding
HIV-1
Transfer RNA
Cells
Amber
Amino acids
Amino Acids
Bearings (structural)
Anticodon
Mutagenesis
Vaccines
Nonsense Codon
Nucleotides
Escherichia coli
Genetic Code
AIDS Vaccines
Genetic Engineering
Terminator Codon

Keywords

  • HIV
  • genetic code
  • quadruplet codon
  • unnatural amino acid
  • virus engineering.

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

Controlling the Replication of a Genomically Recoded HIV-1 with a Functional Quadruplet Codon in Mammalian Cells. / Chen, Yan; Wan, Yanmin; Wang, Nanxi; Yuan, Zhe; Niu, Wei; Li, Qingsheng; Guo, Jiantao.

In: ACS Synthetic Biology, Vol. 7, No. 6, 15.06.2018, p. 1612-1617.

Research output: Contribution to journalArticle

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