Establishing Programmable CRISPR/Cas13b-Mediated Knockdown System in Rice  

作　　者：WANG Shuman ZHANG Linqi GAO Ruiren WEI Guangbo DONG Weiguo XU Jiming WANG Zhiye 

机构地区：[1]State Key Laboratory of Plant Environmental Resilience,College of Life Sciences,Zhejiang University,Hangzhou 310058,China [2]The First Affiliated Hospital,College of Medicine,Zhejiang University,Hangzhou 310058,China

出　　处：《Rice science》2025年第2期217-227,I0043-I0046,共15页水稻科学(英文版)

基　　金：supported by the Natural Science Foundation of Zhejiang Province,China(Grant Nos.LZ22C150002 and LR24C150001);the National Key Research and Development Program of China(Grant Nos.2021YFF1000402 and 2022YFD1401600);the National Natural Science Foundation of China(Grant No.32170262);the Fundamental Research Funds for the Central Universities,China(Grant No.K20240124).

摘　　要：CRISPR-Cas endonucleases mediate prokaryotic adaptive immunity by targeting foreign nucleic acids.CRISPR/Cas13b is a class 2 type VI-B ribonuclease that targets and cleaves single-stranded RNA.It exhibits higher RNA interference activity than Cas13a and Cas13c and causes fewer collateral effects than RxCas13d in mammalian cells.However,a programmable CRISPR/Cas13b-mediated RNA interference system for endogenous transcripts in rice has not yet been established.Here,we developed a CRISPR/Cas13b-mediated system to target endogenous transcripts in rice.Our CRISPR/Cas13b system could inhibit multiple endogenous mRNAs simultaneously.In addition,this system efficiently repressed endogenous long noncoding RNAs with more than 50% inhibition in stable transgenic plants.Furthermore,we found only weak collateral effects of the CRISPR/Cas13b-mediated system at the transcriptome-wide level,and no difference in the agronomic traits of stable transgenic rice in the field.We present a programmable CRISPR/Cas13b-mediated knockdown system for rice,offering a potential biotechnological tool for functional genomics and crop improvement.

关 键 词：CRISPR/Cas13b RNA knockdown system RICE collateral effect inorganic phosphate starvation 

分 类 号：Q78[生物学—分子生物学]