Sequencing of D/L -DNA and XNA by Templated-Synthesis

Article Article scientifique Article Sequencing of D/L -DNA and XNA by Templated-Synthesis eng 2025-02-11 2025-02-19 author Joshi Saurabh Department of Organic Chemistry, CVU, Faculty of Sciences, University of Geneva, 1211 Geneva, Switzerland. 402861 author Romanens Patrick Department of Organic Chemistry, CVU, Faculty of Sciences, University of Geneva, 1211 Geneva, Switzerland. 22629 Département de chimie organique (CHIOR) author Winssinger Nicolas 0000-0003-1636-7766 Department of Organic Chemistry, CVU, Faculty of Sciences, University of Geneva, 1211 Geneva, Switzerland. 769664 Département de chimie organique (CHIOR) 6288-6296 10.1021/jacs.5c00708 39930695 0002-7863 PMC11848921 https://pubs.acs.org/doi/10.1021/jacs.5c00708 Progress in oligonucleotide sequencing has transformed modern biology and medicine. Here we report a fast and efficient enzyme-free primer extension of PNA with reversible chain termination and its application to DNA and XNA sequencing. The approach leverages activated 4-mer PNAs that react in a templated ligation reaction at μM concentrations within minutes. We demonstrate that the fidelity of this enzyme-free primer extension benefits from reactions performed with a mixture of activated PNAs where every 4-mer has its self-complementary 4-mer. The reactions can be performed using the whole repertoire of 4-mers (256 permutations) in a parallelized manner. Using a primer in combination with its -1, -2, and -3 deletion allows for sequencing by MALDI analysis, using the increment in mass for each nucleobase assignment. Given the enzyme-free nature of this sequencing and the achiral nature of PNA, we further demonstrate that the technology can be used to sequence d- or l-DNA as well as LNA and PNA (XNA). Dewey 540 36 Département de chimie organique 141 false 762 Groupe Winssinger Journal of the American Chemical Society 147 7 3f98239e-7635-4590-9c79-9ce32b2bae7c Article (Published version) Public joshi-et-al-2025-sequencing-of-d-l-.pdf 6132102 application/pdf CC BY-4.0 c8d0a9724e0e87f4bb40b67c7b541c86dd47cafe668a39c6114dcf1742f91d55 Vignette Public ja5c00708_0005.png 62111 image/png openAccess Faculté des sciences / Section de chimie et biochimie / Département de chimie organique

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Sequencing of _D/_L -DNA and XNA by Templated-Synthesis	Journal of the American Chemical Society			2025	36	50

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Joshi, Saurabh