Resistance of the CRISPR-Cas13a Gene-Editing System to Potato Spindle Tuber Viroid Infection in Tomato and <i>Nicotiana benthamiana</i>

Gene-editing technology, specifically the CRISPR-Cas13a system, has shown promise in breeding plants resistant to RNA viruses. This system targets RNA and, theoretically, can also combat RNA-based viroids. To test this, the CRISPR-Cas13a system was introduced into tomato plants via transient express...

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Main Authors: Ying Wei Khoo, Qingsong Wang, Shangwu Liu, Binhui Zhan, Tengfei Xu, Wenxia Lv, Guangjing Liu, Shifang Li, Zhixiang Zhang
Format: Article
Language:English
Published: MDPI AG 2024-08-01
Series:Viruses
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Online Access:https://www.mdpi.com/1999-4915/16/9/1401
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author Ying Wei Khoo
Qingsong Wang
Shangwu Liu
Binhui Zhan
Tengfei Xu
Wenxia Lv
Guangjing Liu
Shifang Li
Zhixiang Zhang
author_facet Ying Wei Khoo
Qingsong Wang
Shangwu Liu
Binhui Zhan
Tengfei Xu
Wenxia Lv
Guangjing Liu
Shifang Li
Zhixiang Zhang
author_sort Ying Wei Khoo
collection DOAJ
description Gene-editing technology, specifically the CRISPR-Cas13a system, has shown promise in breeding plants resistant to RNA viruses. This system targets RNA and, theoretically, can also combat RNA-based viroids. To test this, the CRISPR-Cas13a system was introduced into tomato plants via transient expression and into <i>Nicotiana benthamiana</i> through transgenic methods, using CRISPR RNAs (crRNAs) targeting the conserved regions of both sense and antisense genomes of potato spindle tuber viroid (PSTVd). In tomato plants, the expression of CRISPR-Cas13a and crRNAs substantially reduced PSTVd accumulation and alleviated disease symptoms. In transgenic <i>N. benthamiana</i> plants, the PSTVd levels were lower as compared to wild-type plants. Several effective crRNAs targeting the PSTVd genomic RNA were also identified. These results demonstrate that the CRISPR-Cas13a system can effectively target and combat viroid RNAs, despite their compact structures.
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id doaj-art-8ce6578324cb44f4863152d28310f58a
institution OA Journals
issn 1999-4915
language English
publishDate 2024-08-01
publisher MDPI AG
record_format Article
series Viruses
spelling doaj-art-8ce6578324cb44f4863152d28310f58a2025-08-20T01:56:13ZengMDPI AGViruses1999-49152024-08-01169140110.3390/v16091401Resistance of the CRISPR-Cas13a Gene-Editing System to Potato Spindle Tuber Viroid Infection in Tomato and <i>Nicotiana benthamiana</i>Ying Wei Khoo0Qingsong Wang1Shangwu Liu2Binhui Zhan3Tengfei Xu4Wenxia Lv5Guangjing Liu6Shifang Li7Zhixiang Zhang8State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaInstitute of Industrial Crops, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaDepartment of Fruit Science, College of Horticulture, China Agricultural University, Beijing 100193, ChinaInner Mongolia Zhongjia Agricultural Biotechnology Co., Ltd., Ulanqab 011800, ChinaInner Mongolia Zhongjia Agricultural Biotechnology Co., Ltd., Ulanqab 011800, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaGene-editing technology, specifically the CRISPR-Cas13a system, has shown promise in breeding plants resistant to RNA viruses. This system targets RNA and, theoretically, can also combat RNA-based viroids. To test this, the CRISPR-Cas13a system was introduced into tomato plants via transient expression and into <i>Nicotiana benthamiana</i> through transgenic methods, using CRISPR RNAs (crRNAs) targeting the conserved regions of both sense and antisense genomes of potato spindle tuber viroid (PSTVd). In tomato plants, the expression of CRISPR-Cas13a and crRNAs substantially reduced PSTVd accumulation and alleviated disease symptoms. In transgenic <i>N. benthamiana</i> plants, the PSTVd levels were lower as compared to wild-type plants. Several effective crRNAs targeting the PSTVd genomic RNA were also identified. These results demonstrate that the CRISPR-Cas13a system can effectively target and combat viroid RNAs, despite their compact structures.https://www.mdpi.com/1999-4915/16/9/1401tomatoviroidgene editingCas13agenetic engineeringguide RNAs
spellingShingle Ying Wei Khoo
Qingsong Wang
Shangwu Liu
Binhui Zhan
Tengfei Xu
Wenxia Lv
Guangjing Liu
Shifang Li
Zhixiang Zhang
Resistance of the CRISPR-Cas13a Gene-Editing System to Potato Spindle Tuber Viroid Infection in Tomato and <i>Nicotiana benthamiana</i>
Viruses
tomato
viroid
gene editing
Cas13a
genetic engineering
guide RNAs
title Resistance of the CRISPR-Cas13a Gene-Editing System to Potato Spindle Tuber Viroid Infection in Tomato and <i>Nicotiana benthamiana</i>
title_full Resistance of the CRISPR-Cas13a Gene-Editing System to Potato Spindle Tuber Viroid Infection in Tomato and <i>Nicotiana benthamiana</i>
title_fullStr Resistance of the CRISPR-Cas13a Gene-Editing System to Potato Spindle Tuber Viroid Infection in Tomato and <i>Nicotiana benthamiana</i>
title_full_unstemmed Resistance of the CRISPR-Cas13a Gene-Editing System to Potato Spindle Tuber Viroid Infection in Tomato and <i>Nicotiana benthamiana</i>
title_short Resistance of the CRISPR-Cas13a Gene-Editing System to Potato Spindle Tuber Viroid Infection in Tomato and <i>Nicotiana benthamiana</i>
title_sort resistance of the crispr cas13a gene editing system to potato spindle tuber viroid infection in tomato and i nicotiana benthamiana i
topic tomato
viroid
gene editing
Cas13a
genetic engineering
guide RNAs
url https://www.mdpi.com/1999-4915/16/9/1401
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