Photoinduced electron transfer detection method for identifying UGT1A128 microsatellites
During development of a novel detection method for the UDP-glucuronosyl transferase 1A1 (UGT1A1)*28, the fluorescence intensity of a dye conjugated to cytosine (C) at the end of a DNA strand decreased upon hybridization with guanine (G). This phenomenon is referred to as photoinduced electron transf...
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| Veröffentlicht in: | PloS one 2023-08, Vol.18 (8), p.e0289506-e0289506 |
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| creator | Tsuchida, Shirou Himi, Noriaki Miura, Yuuki Kodama, Suzune Shindo, Tsugumi Nakagawa, Koji Aoki, Takashi |
| description | During development of a novel detection method for the UDP-glucuronosyl transferase 1A1 (UGT1A1)*28, the fluorescence intensity of a dye conjugated to cytosine (C) at the end of a DNA strand decreased upon hybridization with guanine (G). This phenomenon is referred to as photoinduced electron transfer (PeT). Using this phenomenon, we devised a method for the naked-eye detection of UGT1A1*28 (thymine-adenine (TA)-repeat polymorphism). Fluorescently labeled single-stranded DNA (ssDNA) oligonucleotides (probes) were designed and hybridized with complementary strand DNAs (target DNAs). Base pair formation at the blunt end between fluorescently labeled C (probe side) and G (target side), induced dramatic fluorescence quenching. Additionally, when the labeled-CG pair formed near the TA-repeat sequence, different TA-repeat numbers were discriminated. However, obtaining enough target DNA for this probe by typical polymerase chain reaction (PCR) was difficult. To enable the practical use of the probe, producing sufficient target DNA remains problematic. |
| doi_str_mv | 10.1371/journal.pone.0289506 |
| format | Article |
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This phenomenon is referred to as photoinduced electron transfer (PeT). Using this phenomenon, we devised a method for the naked-eye detection of UGT1A1*28 (thymine-adenine (TA)-repeat polymorphism). Fluorescently labeled single-stranded DNA (ssDNA) oligonucleotides (probes) were designed and hybridized with complementary strand DNAs (target DNAs). Base pair formation at the blunt end between fluorescently labeled C (probe side) and G (target side), induced dramatic fluorescence quenching. Additionally, when the labeled-CG pair formed near the TA-repeat sequence, different TA-repeat numbers were discriminated. However, obtaining enough target DNA for this probe by typical polymerase chain reaction (PCR) was difficult. To enable the practical use of the probe, producing sufficient target DNA remains problematic.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0289506</identifier><identifier>PMID: 37535593</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adenine ; Analysis ; Biology and Life Sciences ; Cytosine ; Deoxyribonucleic acid ; Digital cameras ; Digital imaging ; DNA ; DNA probes ; Electron transfer ; Electrons ; Fluorescence ; Gene mutations ; Genetic polymorphisms ; Genetic testing ; Hybridization ; Microsatellites ; Nucleotide sequence ; Oligonucleotides ; Pair bond ; Physical sciences ; Polymerase chain reaction ; Polymorphism ; Research and Analysis Methods ; Single-stranded DNA ; Thymine</subject><ispartof>PloS one, 2023-08, Vol.18 (8), p.e0289506-e0289506</ispartof><rights>Copyright: © 2023 Tsuchida et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</rights><rights>COPYRIGHT 2023 Public Library of Science</rights><rights>2023 Tsuchida et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2023 Tsuchida et al 2023 Tsuchida et al</rights><rights>2023 Tsuchida et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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This phenomenon is referred to as photoinduced electron transfer (PeT). Using this phenomenon, we devised a method for the naked-eye detection of UGT1A1*28 (thymine-adenine (TA)-repeat polymorphism). Fluorescently labeled single-stranded DNA (ssDNA) oligonucleotides (probes) were designed and hybridized with complementary strand DNAs (target DNAs). Base pair formation at the blunt end between fluorescently labeled C (probe side) and G (target side), induced dramatic fluorescence quenching. Additionally, when the labeled-CG pair formed near the TA-repeat sequence, different TA-repeat numbers were discriminated. However, obtaining enough target DNA for this probe by typical polymerase chain reaction (PCR) was difficult. To enable the practical use of the probe, producing sufficient target DNA remains problematic.</description><subject>Adenine</subject><subject>Analysis</subject><subject>Biology and Life Sciences</subject><subject>Cytosine</subject><subject>Deoxyribonucleic acid</subject><subject>Digital cameras</subject><subject>Digital imaging</subject><subject>DNA</subject><subject>DNA probes</subject><subject>Electron transfer</subject><subject>Electrons</subject><subject>Fluorescence</subject><subject>Gene mutations</subject><subject>Genetic polymorphisms</subject><subject>Genetic testing</subject><subject>Hybridization</subject><subject>Microsatellites</subject><subject>Nucleotide sequence</subject><subject>Oligonucleotides</subject><subject>Pair bond</subject><subject>Physical sciences</subject><subject>Polymerase chain reaction</subject><subject>Polymorphism</subject><subject>Research and Analysis Methods</subject><subject>Single-stranded 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This phenomenon is referred to as photoinduced electron transfer (PeT). Using this phenomenon, we devised a method for the naked-eye detection of UGT1A1*28 (thymine-adenine (TA)-repeat polymorphism). Fluorescently labeled single-stranded DNA (ssDNA) oligonucleotides (probes) were designed and hybridized with complementary strand DNAs (target DNAs). Base pair formation at the blunt end between fluorescently labeled C (probe side) and G (target side), induced dramatic fluorescence quenching. Additionally, when the labeled-CG pair formed near the TA-repeat sequence, different TA-repeat numbers were discriminated. However, obtaining enough target DNA for this probe by typical polymerase chain reaction (PCR) was difficult. To enable the practical use of the probe, producing sufficient target DNA remains problematic.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>37535593</pmid><doi>10.1371/journal.pone.0289506</doi><tpages>e0289506</tpages><orcidid>https://orcid.org/0000-0002-1625-7770</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Adenine Analysis Biology and Life Sciences Cytosine Deoxyribonucleic acid Digital cameras Digital imaging DNA DNA probes Electron transfer Electrons Fluorescence Gene mutations Genetic polymorphisms Genetic testing Hybridization Microsatellites Nucleotide sequence Oligonucleotides Pair bond Physical sciences Polymerase chain reaction Polymorphism Research and Analysis Methods Single-stranded DNA Thymine |
| title | Photoinduced electron transfer detection method for identifying UGT1A128 microsatellites |
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