Enzymatic Synthesis Using Polymerases of Modified Nucleic Acids and Genes

Eremeeva, Elena; Herdewijn, Piet

doi:10.1002/9783527812103.ch7

“…Similarly, enzymatic XNA synthesis may facilitate production of XNA oligonucleotides. XNAs with chemically conservative modifications are accepted as substrates by natural polymerases, while engineering of polymerases to accept a broader range of XNA substrates has also met with success [72,73]. Indeed, some polymerases have proven highly adaptable to new substrates.…”

Section: Polymerase Synthesis Of Xnasmentioning

confidence: 99%

Modified nucleic acids: replication, evolution, and next-generation therapeutics

Duffy

¹

,

Arangundy-Franklin

²

,

Holliger

³

2020

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Modified nucleic acids, also called xeno nucleic acids (XNAs), offer a variety of advantages for biotechnological applications and address some of the limitations of first-generation nucleic acid therapeutics. Indeed, several therapeutics based on modified nucleic acids have recently been approved and many more are under clinical evaluation. XNAs can provide increased biostability and furthermore are now increasingly amenable to in vitro evolution, accelerating lead discovery. Here, we review the most recent discoveries in this dynamic field with a focus on progress in the enzymatic replication and functional exploration of XNAs.

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The Network of Replication, Transcription, and Reverse Transcription of a Synthetic Genetic Cassette

Yang

¹

,

Eremeeva

²

,

Abramov

³

et al. 2020

Angewandte Chemie

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1

0

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Synthetic nucleic acids, with four non‐canonical nucleobases, can function as genetic materials. A comprehensive analysis of PCR amplification, transcription, reverse transcription, and cloning was done to screen for alternative genetic monomers. A small library of six modified nucleobases was selected: the modified 2′‐deoxyribonucleoside (dZTPs) and ribonucleoside (rZTPs) triphosphates of 7‐deaza‐adenine, 5‐chlorouracil, 7‐deaza‐guanine or inosine together with 5‐fluorocytosine or 5‐bromocytosine. The fragments composed of one to four modified nucleotides (denoted as DZA) have been successfully recognized and transcribed to natural or modified RNA (denoted as RZA) by T7 RNA polymerase. The fully modified RZA fragment could be reverse transcribed and then amplified in the presence of various dZTPs. Noticeably, modified fragments could function as genetic templates in vivo by encoding the 678 base pair gene of a fluorescent protein in bacteria. These results demonstrate the existence of a fully simulated genetic circuit that uses synthetic materials.

show abstract

The Network of Replication, Transcription, and Reverse Transcription of a Synthetic Genetic Cassette

Yang

¹

,

Eremeeva

²

,

Abramov

³

et al. 2020

Self Cite

View full text Add to dashboard Cite

Synthetic nucleic acids, with four non‐canonical nucleobases, can function as genetic materials. A comprehensive analysis of PCR amplification, transcription, reverse transcription, and cloning was done to screen for alternative genetic monomers. A small library of six modified nucleobases was selected: the modified 2′‐deoxyribonucleoside (dZTPs) and ribonucleoside (rZTPs) triphosphates of 7‐deaza‐adenine, 5‐chlorouracil, 7‐deaza‐guanine or inosine together with 5‐fluorocytosine or 5‐bromocytosine. The fragments composed of one to four modified nucleotides (denoted as DZA) have been successfully recognized and transcribed to natural or modified RNA (denoted as RZA) by T7 RNA polymerase. The fully modified RZA fragment could be reverse transcribed and then amplified in the presence of various dZTPs. Noticeably, modified fragments could function as genetic templates in vivo by encoding the 678 base pair gene of a fluorescent protein in bacteria. These results demonstrate the existence of a fully simulated genetic circuit that uses synthetic materials.

show abstract

Enzymatic Synthesis Using Polymerases of Modified Nucleic Acids and Genes

Cited by 4 publications

References 136 publications

Modified nucleic acids: replication, evolution, and next-generation therapeutics

Modified nucleic acids: replication, evolution, and next-generation therapeutics

The Network of Replication, Transcription, and Reverse Transcription of a Synthetic Genetic Cassette

The Network of Replication, Transcription, and Reverse Transcription of a Synthetic Genetic Cassette

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