Efficient synthesis and replication of diverse sequence libraries composed of biostable nucleic acid analogues

John R.D. Hervey; Niklas Freund; Gillian Houlihan; Gurpreet Dhaliwal; Philipp Holliger; Alexander I. Taylor

doi:10.1039/d2cb00035k

Efficient synthesis and replication of diverse sequence libraries composed of biostable nucleic acid analogues

John R.D. Hervey, Niklas Freund, Gillian Houlihan, Gurpreet Dhaliwal, Philipp Holliger^*, Alexander I. Taylor^*

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

11 Citations (Scopus)

Abstract

Functional nucleic acids can be evolved in vitro using cycles of selection and amplification, starting from diverse-sequence libraries, which are typically restricted to natural or partially-modified polymer chemistries. Here, we describe the efficient DNA-templated synthesis and reverse transcription of libraries entirely composed of serum nuclease resistant alternative nucleic acid chemistries validated in nucleic acid therapeutics; locked nucleic acid (LNA), 2′-O-methyl-RNA (2′OMe-RNA), or mixtures of the two. We evaluate yield and diversity of synthesised libraries and measure the aggregate error rate of a selection cycle. We find that in addition to pure 2′-O-methyl-RNA and LNA, several 2′OMe-RNA/LNA blends seem suitable and promising for discovery of biostable functional nucleic acids for biomedical applications.

Original language	English
Pages (from-to)	1209-1215
Number of pages	7
Journal	RSC Chemical Biology
Volume	3
Issue number	10
DOIs	https://doi.org/10.1039/d2cb00035k
Publication status	Published - 30 Aug 2022

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AU - Freund, Niklas

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AU - Holliger, Philipp

AU - Taylor, Alexander I.

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N2 - Functional nucleic acids can be evolved in vitro using cycles of selection and amplification, starting from diverse-sequence libraries, which are typically restricted to natural or partially-modified polymer chemistries. Here, we describe the efficient DNA-templated synthesis and reverse transcription of libraries entirely composed of serum nuclease resistant alternative nucleic acid chemistries validated in nucleic acid therapeutics; locked nucleic acid (LNA), 2′-O-methyl-RNA (2′OMe-RNA), or mixtures of the two. We evaluate yield and diversity of synthesised libraries and measure the aggregate error rate of a selection cycle. We find that in addition to pure 2′-O-methyl-RNA and LNA, several 2′OMe-RNA/LNA blends seem suitable and promising for discovery of biostable functional nucleic acids for biomedical applications.

AB - Functional nucleic acids can be evolved in vitro using cycles of selection and amplification, starting from diverse-sequence libraries, which are typically restricted to natural or partially-modified polymer chemistries. Here, we describe the efficient DNA-templated synthesis and reverse transcription of libraries entirely composed of serum nuclease resistant alternative nucleic acid chemistries validated in nucleic acid therapeutics; locked nucleic acid (LNA), 2′-O-methyl-RNA (2′OMe-RNA), or mixtures of the two. We evaluate yield and diversity of synthesised libraries and measure the aggregate error rate of a selection cycle. We find that in addition to pure 2′-O-methyl-RNA and LNA, several 2′OMe-RNA/LNA blends seem suitable and promising for discovery of biostable functional nucleic acids for biomedical applications.

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Efficient synthesis and replication of diverse sequence libraries composed of biostable nucleic acid analogues

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