vestichИзвестия Национальной академии наук Беларуси. Серия химических наукProceedings of the National Academy of Sciences of Belarus, Chemical Series1561-83312524-2342The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1561-8331-2022-58-4-398-406vestich-764Research ArticleБИООРГАНИЧЕСКАЯ ХИМИЯBIOORGANIC CHEMISTRYОптимизация технологии получения направляющих РНК на планшетном автоматическом синтезатореOptimization of the technology for obtaining guide RNA using plate automatic synthesizerУлащикЕ. А.UlashchikE. A.

Улащик Егор Александрович – научный сотрудник.

Ул. Сурганова, 13, 220072, Минск

Ulashchik Egor A. – Researcher.

13, Surganov Str., 220072, Minsk

e.ulashchik@gmail.comАхламёнокТ. П.AkhlamionokT. P.

Ахламёнок Татьяна Петровна – младший научный сотрудник.

Ул. Сурганова, 13, 220072, Минск

Akhlamionok Tatsiana P. – Junior Researcher.

13, Surganov Str., 220072, Minsk

t.akhlamionak@gmail.comБорищикП. Ю.BaryshchykP. Y.

Борищик Полина Юрьевна – младший научный сотрудник.

Ул. Сурганова, 13, 220072, Минск

Baryshchyk Palina Y. – Junior Researcher.

13, Surganov Str., 220072, Minsk

contact.polina@yandex.byШаркоО. Л.SharkoO. L.

Шарко Ольга Леонидовна – кандидат химических наук, ведущий научный сотрудник.

Ул. Сурганова, 13, 220072, Минск

Sharko Olga L. – Ph. D. (Chemistry), Leading Researcher.

13, Surganov Str., 220072, Minsk

sharko@ifoch.bas-net.byШманайВ. В.ShmanaiV. V.

Шманай Вадим Владимирович – кандидат химических наук, заведующий лабораторией.

Ул. Сурганова, 13, 220072, Минск

Shmanai Vadim V. – Ph. D. (Chemistry), Head of the Laboratory.

13, Surganov Str., 220072, Minsk

shmanai@ifoch.bas-net.byИнститут физико-органической химии, Национальная академия наук БеларусиShmanai Institute of Physical Organic Chemistry, National Academy of Sciences of BelarusИнститут физико-органической химии, Национальная академия наук БеларусиInstitute of Physical Organic Chemistry, National Academy of Sciences of Belarus202229112022584398406Copyright © Улащик Е.А., Ахламёнок Т.П., Борищик П.Ю., Шарко О.Л., Шманай В.В., 20222022Улащик Е.А., Ахламёнок Т.П., Борищик П.Ю., Шарко О.Л., Шманай В.В.Ulashchik E.A., Akhlamionok T.P., Baryshchyk P.Y., Sharko O.L., Shmanai V.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestichem.belnauka.by/jour/article/view/764

Разработан протокол синтеза и подобраны подходящие реагенты для получения направляющих РНК-олигонуклеотидов для системы CRISPR/Cas на автоматическом синтезаторе ASM-2000 в масштабе 500 нмоль. Отработаны методики выделения, очистки и аналитического контроля синтетических РНК-олигонуклеотидов. С использованием улучшенной технологии получены олигонуклеотиды, являющиеся направляющими РНК для системы CRISPR Cas12a.

A synthetic protocol was developed and optimal reagents have been selected for obtaining guide RNA oligonucleotides for the CRISPR/Cas system using ASM-2000 automatic synthesizer in 500 nmol scale. Methods for the isolation, purification and analytical control of synthetic RNA oligonucleotides have been developed. The improved technology has been used for preparation of guide RNAs for the CRISPR Cas12a system.

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The authors declare that there are no conflicts of interest present.