Design and Synthesis of a Photocleavable Fluorescent Nucleotide 3‘-O-Allyl-dGTP-PC-Bodipy-FL-510 as a Reversible Terminator for DNA Sequencing by Synthesis

DNA sequencing by synthesis (SBS) using reversible fluorescent nucleotide terminators is potentially an efficient approach to address the limitations of current DNA sequencing techniques. Here, we report the design and synthesis of a 3‘-O-allyl photocleavable fluorescent nucleotide analogue, 3‘-O-al...

Full description

Saved in:
Bibliographic Details
Published in:Journal of organic chemistry 2006-04, Vol.71 (8), p.3248-3252
Main Authors: Meng, Qinglin, Kim, Dae Hyun, Bai, Xiaopeng, Bi, Lanrong, Turro, Nicholas J, Ju, Jingyue
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Base Sequence
Biological and medical sciences
Carbohydrates. Nucleosides and nucleotides
Chemistry
Deoxyguanine Nucleotides - chemical synthesis
Deoxyguanine Nucleotides - chemistry
Deoxyguanine Nucleotides - metabolism
Dna, deoxyribonucleoproteins
DNA-Directed DNA Polymerase - metabolism
Exact sciences and technology
Fluorescent Dyes - chemical synthesis
Fluorescent Dyes - chemistry
Fluorescent Dyes - metabolism
Fundamental and applied biological sciences. Psychology
Nucleic acids
Nucleosides, nucleotides and oligonucleotides
Organic chemistry
Photochemistry
Preparations and properties
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:DNA sequencing by synthesis (SBS) using reversible fluorescent nucleotide terminators is potentially an efficient approach to address the limitations of current DNA sequencing techniques. Here, we report the design and synthesis of a 3‘-O-allyl photocleavable fluorescent nucleotide analogue, 3‘-O-allyl-dGTP-PC-Bodipy-FL-510, as a reversible terminator for SBS. The nucleotide is efficiently incorporated by DNA polymerase into a growing DNA strand to terminate the polymerase reaction. After that, the fluorophore is photocleaved quantitatively by irradiation at 355 nm, and the allyl group is rapidly and efficiently removed by using a Pd-catalyzed reaction under DNA-compatible conditions to regenerate a free 3‘-OH group to reinitiate the polymerase reaction. Two cycles of such steps were successfully demonstrated to sequence a homopolymeric region of a DNA template, facilitating the development of SBS as a viable approach for high-throughput DNA sequencing.
ISSN:0022-3263
1520-6904
DOI:10.1021/jo060300k