Efficient deletion of multiple circle RNA loci by CRISPR‐Cas9 reveals Os06circ02797 as a putative sponge for OsMIR408 in rice

• DOI: 10.1111/pbi.13544
• Publication Date: 2021-01-15T15:18:46Z
• Authors: Jianping Zhou; Mingzhu Yuan; Yuxin Zhao; Quan Quan; Dong Yu; Han Yang; Xu Tang; Xuhui Xin; Guangze Cai; Qian Qian; Yiping Qi; Yong Zhang
• Keywords: Gene Editing; 0301 basic medicine; 2. Zero hunger; 03 medical and health sciences; Chlorophyll A; Oryza; RNA, Circular; CRISPR-Cas Systems; Plants, Genetically Modified; Research Articles

Summary CRISPR‐Cas9 is an emerging genome editing tool for reverse genetics in plants. However, its application functional study of non‐coding RNAs plants still at infancy. Despite being a major class RNAs, the biological roles circle (circRNAs) remain largely unknown Previous plant circRNA studies have focused on identification and annotation putative circRNAs, with their functions uninvestigated by genetic approaches. Here, we applied multiplexed strategy to efficiently acquire individual null mutants four circRNAs rice. We showed each these rice loci ( Os02circ25329 , Os06circ02797 Os03circ00204 Os05circ02465 ) can be deleted 10% or higher efficiency both protoplasts stable transgenic T0 lines. Such high deletion enabled generation allele without transgene T1 generation. Characterization reveals circRNAs’ participation salt stress response during seed germination particular mutant tolerance. Notably, seedlings rapid growth phenotype after containing chlorophyll A/B content. Further molecular computational analyses suggested circRNA–miRNA–mRNA regulatory network where bind sequester OsMIR408 important conserved microRNA This not only presents evidence first time that certain may serve as sponges negatively regulate miRNAs, phenomenon previously demonstrated mammalian cells, but also provides insights improving agronomic traits through gene crops.