Table 1.
Summary presentation of the efforts done using multiplex genome editing for revolutionizing plant biology and crop improvement programs.
| MGE application | Target genes/QTLs | Crop | Objective | References |
|---|---|---|---|---|
| Plant biology | ||||
| Knock-out of multiple genes | ||||
| Two targets at ssDNA tomato yellow leaf curl virus (TYLCV) viruses | Tomato | Virus biological control in plants | Ali et al. (2015) | |
| OsGW2, OsGW5, and OsTGW6 | Rice | Larger grain size | Xu et al. (2016) | |
| RAS-PDS1 and RAS-PDS2 | Banana | Demonstration of gene knockout in banana | Kaur et al. (2018) | |
| OsSWEET11 and OsSWEET14 | Rice | Broad-spectrum disease resistance in rice | Xu et al. (2019) | |
| TMS5, Pi21, and Xa13 | Rice | Hybrid rice production and disease resistance | Li S. et al. (2019) | |
| Engineering efficient plant metabolic pathways | ||||
| OsGSTU, OsMRP15, and OsAnP | Rice | Anthocyanin synthesis pathway | Ma et al. (2015) | |
| Three different sites in the OsWaxy | Rice | Decreasing amylose content | Ma et al. (2015) | |
| Two sites in slyPDS | Tomato | Generating photobleached phenotype | Li R. et al. (2018) | |
| GABA-TP1, GABA-TP2, GABA-TP3, CAT9, and SSADH | Tomato | Metabolic engineering and manipulation of the gamma-aminobutyric acid (GABA) | Li R. et al. (2018) | |
| SGR1, LCY-E, Blc, LCY-B1, and LCY-B2 | Tomato | Increase the accumulation of lycopene | Li X. et al. (2018) | |
| GmF3H1, GmF3H2, and GmFNSII-1 | Soybean | Increasing the isoflavone content | Zhang et al. (2020) | |
| Transcriptional regulation | ||||
| Three different sites in the promoter of production of Anthocyanin Pigment1 gene (AtPAP1, encoding a transcription factor) and miR319 (encoding a microRNA) | Arabidopsis | Transcription gene activation | Lowder et al. (2015) | |
| Promoter regions of the WRKY30, RLP23, and CDG1 genes | Arabidopsis | Simultaneous activation of their transcription | Li et al. (2017) | |
| Two sites at locule number (lc) QTL + eight gRNAs targeting the promoters of SlCLV3, S and SP | Tomato | Generating desirable/beneficial regulatory variants | Rodríguez-Leal et al. (2017) | |
| OsWOX11 and OsYUC1 | Rice | Transcription gene activation | Gong et al. (2020) | |
| Chromosomal segment restructuring | ||||
| Two targets flanking a 245 kb segment | Rice | Chromosomal segment deletions | Zhou et al. (2014) | |
| Two targets flanking 1.8 kb segment | Tobacco | Chromosomal segment deletions and inversion | Gao et al. (2015) | |
| Two targets flanking a 100 bp segment | Arabidopsis | Chromosomal segment deletions | Ordon et al. (2017) | |
| 6 targets at a 58 kb region | Medicago | Chromosomal segment deletions | Cermák et al. (2017) | |
| Different genomic regions ranging from 1.7 to 13 kb | Arabidopsis | Chromosomal segment deletions | Wu et al. (2018) | |
| Two targets flanking an 18 (kb) segment | Arabidopsis | Chromosomal inversion | Schmidt et al. (2019) | |
| Multiplex base alterations | ||||
| Three sgRNAs targeting ALS and FTIP1e genes | Rice | Herbicide resistance | Shimatani et al. (2017) | |
| OsACC-T1, OsALS-T1, OsCDC48-T3, OsDEP1-T1, OsDEP1-T2, and OsNRT1.1B-T1. | Rice | Herbicide resistance | Li C. et al. (2018) | |
| AtALS, AtPDS, AtFT, and AtLFY genes and BnALS and BnPDS | Arabidopsis and Rapeseed | System verification | Kang et al. (2018) | |
| Two targets StGBSSI gene | Potato | impaired amylose biosynthesis in potato | Veillet et al. (2019) | |
| ZmALS1 and ZmALS2 | Maize | Herbicide resistance | Li Y. et al. (2019) | |
| Four targets at OsWAXY, OsCDC48, and OsSNB genes | Rice | System optimization | Wang F. et al. (2020) | |
| BnALS1 and BnALS3 | Oilseed rape | Herbicide resistance | Wu et al. (2020) | |
| Multiple sites at OsALS1 gene | Rice | Facilitating the directed evolution of plant | Kuang et al. (2020) | |
| Multiple loci at OsACC | Rice | Herbicide resistance | Liu et al. (2020) | |
| Multiple targets trials | Rice | Prime genome editing optimization | Lin et al. (2020) | |
| Crop improvement | ||||
| Generating diversity and crop domestication | ||||
| Two sites at locule number (Lc) QTL + eight gRNAs targeting the promoters of SlCLV3, S and SP. | Tomato | Wild relative domestication | Rodríguez-Leal et al. (2017) | |
| Six targets at SP, SP5G, SlCLV3 and SlWUS | Tomato | Wild relative domestication | Li T. et al. (2018) | |
| Six targets at SELF PRUNING, OVATE, FRUIT WEIGHT 2.2, LYCOPENE BETA-CYCLASE, MULTIFLORA and FASCIATED/YABBY genes | Tomato | Domestication of wild Solanum pimpinellifolium. | Zsögön et al. (2018) | |
| GS3, GW2, and GN1A | Rice | Wild relative domestication (Oryza glaberrima) | Lacchini et al. (2020) | |
| Multiple | Rice | Wild relative domestication (O. alta) | Yu et al. (2021) | |
| Increasing crop yield potential | ||||
| OsGW2, OsGW5, and OsTGW6 | Rice | Larger grain size | Xu et al. (2016) | |
| OsGS3, OsGW2, and OsGn1a | Rice | Larger grain size | Zhou et al. (2019) | |
| GS3, GW2, and GN1A | Rice | Increasing grain size | Lacchini et al. (2020) | |
| Improving crop product quality | ||||
| 4 alleles of GBSS | Potato | Produce waxy potato | Andersson et al. (2017) | |
| OVATE, FRUIT WEIGHT 2.2 and LYCOPENE BETA CYCLASE | Tomato | Fruit shape | Zsögön et al. (2018) | |
| Several sites at St16DOX | Potato | Reducing steroidal glycoalkaloids (SGAs) | Nakayasu et al. (2018) | |
| Two sites at α-gliadin gene | Wheat | Produce low-gluten wheat | Sánchez-León et al. (2018) | |
| OsGS3, OsGW2, and OsGn1a | Rice | Grain shape | Lacchini et al. (2020) | |
| OsGS3, OsGW2, and OsGn1a | Rice | Grain shape | Lacchini et al. (2020) | |
| Enhancing crop resistance/tolerance to biotic/abiotic stress | ||||
| DIPM-1, DIPM-2, and DIPM-4 | Apple | Disease resistance | Malnoy et al. (2016) | |
| OsSWEET11 and OsSWEET14 | Rice | Generating broad spectrum disease resistance | Xu et al. (2019) | |
| SWEET11, SWEET13 and SWEET14 | Rice | Generating broad spectrum disease resistance | Oliva et al. (2019) | |
| TMS5, Pi21, and Xa13 | Rice | Disease resistance and yield enhancement | Li S. et al. (2019) | |