Nature communications · 2022
Engineered Cas12i2 is a versatile high-efficiency platform for therapeutic genome editing.
СтатьяНазвание и аннотация: Перевод в очереди
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
McGaw Colin, Garrity Anthony J, Munoz Gabrielle Z, Haswell Jeffrey R, Sengupta Sejuti, Keston-Smith Elise, Hunnewell Pratyusha, Ornstein Alexa, Bose Mishti, Wessells Quinton, Jakimo Noah, Yan Paul, Zhang Huaibin, Alfonse Lauren E, Ziblat Roy, Carte Jason M, Lu Wei-Cheng, Cerchione Derek, Hilbert Brendan, Sothiselvam Shanmugapriya, Yan Winston X, Cheng David R, Scott David A, DiTommaso Tia, Chong Shaorong
Аннотация
The CRISPR-Cas type V-I is a family of Cas12i-containing programmable nuclease systems guided by a short crRNA without requirement for a tracrRNA. Here we present an engineered Type V-I CRISPR system (Cas12i), ABR-001, which utilizes a tracr-less guide RNA. The compact Cas12i effector is capable of self-processing pre-crRNA and cleaving dsDNA targets, which facilitates versatile delivery options and multiplexing, respectively. We apply an unbiased mutational scanning approach to enhance initially low editing activity of Cas12i2. The engineered variant, ABR-001, exhibits broad genome editing capability in human cell lines, primary T cells, and CD34+ hematopoietic stem and progenitor cells, with both robust efficiency and high specificity. In addition, ABR-001 achieves a high level of genome editing when delivered via AAV vector to HEK293T cells. This work establishes ABR-001 as a versatile, specific, and high-performance platform for ex vivo and in vivo gene therapy.