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Phosphoramidate Azole Oligonucleotides for Single Nucleotide Polymorphism Detection by PCR Научная публикация

Журнал International Journal of Molecular Sciences
, E-ISSN: 1422-0067
Вых. Данные Год: 2024, Том: 25, Номер: 1, Номер статьи : 617, Страниц : DOI: 10.3390/ijms25010617
Ключевые слова PCR; allele-specific PCR; modified oligonucleotides; phosphoramide benzoazole oligonucleotides; PABAO; mutation detection; single nucleotide polymorphism detection; SNP; KRAS mutations
Авторы Chubarov Alexey S. 1 , Baranovskaya Elizaveta E. 1 , Oscorbin Igor P. 1 , Yushin Ivan I. 1 , Filipenko Maxim L. 1 , Pyshnyi Dmitrii V. 1 , Vasilyeva Svetlana V. 1 , Lomzov Alexander A. 1
Организации
1 Institute of Chemical Biology and Fundamental Medicine, SB RAS, 8 Lavrentiev Avenue, 630090 Novosibirsk, Russia

Информация о финансировании (1)

1 Российский научный фонд РНФ №23-14-00358

Реферат: Detection of the Kirsten rat sarcoma gene (KRAS) mutational status is an important factor for the treatment of various malignancies. The most common KRAS-activating mutations are caused by single-nucleotide mutations, which are usually determined by using PCR, using allele-specific DNA primers. Oligonucleotide primers with uncharged or partially charged internucleotide phosphate modification have proved their ability to increase the sensitivity and specificity of various single nucleotide mutation detection. To enhance the specificity of single nucleotide mutation detection, the novel oligonucleotides with four types of uncharged and partially charged internucleotide phosphates modification, phosphoramide benzoazole (PABA) oligonucleotides (PABAO), was used to prove the concept on the KRAS mutation model. The molecular effects of different types of site-specific PABA modification in a primer or a template on a synthesis of full-length elongation product and PCR efficiency were evaluated. The allele-specific PCR (AS-PCR) on plasmid templates showed a significant increase in analysis specificity without changes in Cq values compared with unmodified primer. PABA modification is a universal mismatch-like disturbance, which can be used for single nucleotide polymorphism discrimination for various applications. The molecular insights of the PABA site-specific modification in a primer and a template affect PCR, structural features of four types of PABAO in connection with AS-PCR results, and improvements of AS-PCR specificity support the further design of novel PCR platforms for various biological targets testing.
Библиографическая ссылка: Chubarov A.S. , Baranovskaya E.E. , Oscorbin I.P. , Yushin I.I. , Filipenko M.L. , Pyshnyi D.V. , Vasilyeva S.V. , Lomzov A.A.
Phosphoramidate Azole Oligonucleotides for Single Nucleotide Polymorphism Detection by PCR
International Journal of Molecular Sciences. 2024. V.25. N1. 617 . DOI: 10.3390/ijms25010617 WOS Scopus РИНЦ OpenAlex
Даты:
Опубликована в печати: 3 янв. 2024 г.
Идентификаторы БД:
Web of science: WOS:001141532200001
Scopus: 2-s2.0-85181165855
РИНЦ: 65729487
OpenAlex: W4390544228
Цитирование в БД:
БД Цитирований
OpenAlex 5
Scopus 1
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