1College of Food Science & Nutritional Engineering, China Agricultural University, Beijing, 100083, China 2State Key Laboratory of Rice Biology and Breeding, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310006, China *Corresponding author. E-mail: firstname.lastname@example.org
Received: 02 Apr 2023 Accepted: 14 Aug 2023 Published online: 22 Aug 2023
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is a powerful and versatile gene editing system that has been extensively utilized in various animals and plants, which holds enormous potential and value for scientific research and breeding. However, single-targeted CRISPR can only induce a few base deletions, insertions, or substitution. Ideally, these mutations result in premature termination of the protein encoded by the target gene, leading to a loss of function . Nevertheless, in some cases, these mutations can produce truncated proteins whose still have function . Moreover, when applied to long non-coding RNAs (lncRNAs), which rely on their secondary structures to function, single base mutations may not alter their structures so that they lose their function. Such imprecise genome editing models hinder the accurate identification of gene function. In crops with high gene editing efficiency, such as rice and soybean, dual sgRNAs can induce large fragment deletions on target genes, thereby avoiding these limitations [3, 4]. However, obtaining large fragment deletions through dual sgRNAs is inefficient or rarely reported in most horticultural crops, such as tomato . Hence, efficient technologies for large DNA fragment deletions in horticultural crops are urgently needed for research and breeding purposes.