Two HEPN domains dictate CRISPR RNA maturation and target cleavage in Cas13d
Journal
Nature Communications
Journal Volume
10
Journal Issue
1
Date Issued
2019
Author(s)
Zhang, Bo
Ye, Yangmiao
Ye, Weiwei
Perčulija, Vanja
Jiang, Han
Chen, Yiyang
Li, Yu
Chen, Jing
Lin, Jinying
Wang, Siqi
Chen, Qi
Ouyang, Songying
Abstract
ions aid in stabilizing the conformation of the crRNA repeat region. Sequestration of divalent metal ions does not alter pre-crRNA processing, but abolishes target cleavage by UrCas13d. Notably, the pre-crRNA processing is executed by the HEPN-2 domain. Furthermore, both the structure and sequence of the nucleotides U(-8)-C(-1) within the repeat region are indispensable for target cleavage, and are specifically recognized by UrCas13d. Moreover, correct base pairings within two separate spacer regions (an internal and a 3'-end region) are essential for target cleavage. These findings provide a framework for the development of Cas13d into a tool for a wide range of applications.
SDGs
Type
journal article