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Targeted Modification of Mammalian DNA by a Novel Type V Cas12a Endonuclease from Ruminococcus bromiiстатья Исследовательская статья
Статья опубликована в высокорейтинговом журнале
Статья опубликована в журнале из списка Web of Science и/или Scopus- Авторы: Vasilev Ruslan, Gunitseva Natalia, Shebanova Regina, Korzhenkov Aleksei, Vlaskina Anna, Evteeva Marta, Polushkina Irina, Nikitchina Natalia, Toshchakov Stepan, Kamenski Piotr, Patrushev Maxim, Mazunin Ilya
- Журнал: International Journal of Molecular Sciences
- Том: 23
- Номер: 16
- Год издания: 2022
- Издательство: MDPI
- Местоположение издательства: Basel, Switzerland
- Номер статьи: 9289
- DOI: 10.3390/ijms23169289
- Аннотация: Type V Cas12a nucleases are DNA editors working in a wide temperature range and using expanded protospacer-adjacent motifs (PAMs). Though they are widely used, there is still a demand for discovering new ones. Here, we demonstrate a novel ortholog from Ruminococcus bromii sp. entitled RbCas12a, which is able to efficiently cleave target DNA templates, using the particularly high accessibility of PAM 5′-YYN and a relatively wide temperature range from 20 °C to 42 °C. In comparison to Acidaminococcus sp. (AsCas12a) nuclease, RbCas12a is capable of processing DNA more efficiently, and can be active upon being charged by spacer-only RNA at lower concentrations in vitro. We show that the human-optimized RbCas12a nuclease is also active in mammalian cells, and can be applied for efficient deletion incorporation into the human genome. Given the advantageous properties of RbCas12a, this enzyme shows potential for clinical and biotechnological applications within the field of genome editing.
- Добавил в систему: Каменский Петр Андреевич