TABLE 2.

Applications of CRISPR/Cas9 system in cereal crops.

Crop	Type of study	Targeted gene	Method	sgRNA promoter	Cas9 promoter	Editing efficiency (%)	References
Wheat (Triticum aestivum)	Genome editing	Tainox and Tapds	Agrobacterium-mediated	CaMV 35S	CaMV 35S	11–12	Upadhyay et al. (2013)
	Genome editing	TaGLW7, TaGW8, TaGW2, and TaCKX2-1	Agrobacterium-mediated	TaU6.1, TaU6.2, TaU6.3, and TaU6.5	ZmUbi	64.3	Zhang et al. (2019a)
	Genome editing	TaGASR7A1, TaGASR7 B1, and TaGASR7D1	Biolistic	TaU6	CaMV 35S	5.2	Hamada et al. (2018)
	Knockout	TdGASR7	Biolistic	TaU6	2×CaMV 35S	1.8	Liang et al. (2018)
	Knockout	TaGW2B1, TaGW2D1, and TaGW2A1	Protoplast transfection using RNP	__	__	0–4.4	Liang et al. (2017)
	Gene editing	TaABCC6, TaNFXL1, and TansLTP	Protoplast transformation	TaU6	CaMV 35S	NA	Cui et al. (2019)
	knockout	GW2-B, PinB-D, and ASN2-A	Protoplast transformation using RNP	__	__	0–36	Brandt et al. (2020)
	Genome editing through transient expression	TaGASR7A1, TaGASR7B1, and TaGASR7D1	Particle bombardment	TaU6	Ubi	1.1–5	Zhang et al. (2016)
	Site-directed mutagenesis	TaMLOD1, TaMLOA1, and TaMLOB1	Particle bombardment	U6	Ubi1	3.4–6	Wang et al. (2014)
	Site-directed mutagenesis	LOX2	Protoplast transformation	TaU6	2×CaMV 35S	∼1	Shan et al. (2014)
	Functional genomics	TaPDS	Agrobacterium- mediated	TaU6	ZmUbi	11–17	Howells et al. (2018)
Sorghum (Sorghum bicolor)	Gene editing	TaLox2 and TaUbiL1	Electroporation	TaU6	Ubi1	2.2	Bhowmik et al. (2018)
	Gene editing	CAD and PDS	Particle bombardment	U3	ZmUbi	NA	Liu et al. (2020)
	Gene editing	K1c	Agrobacterium-mediated	TaU3	ZmUbi	14.1–78.3	Li et al. (2018)
	Knockout	SbFT and SbGA2ox5	Agrobacterium-mediated	U6P.1 and U6P.2	ZmUbi	33.3–83.3	Char and Yang. (2019)
	Targeted mutagenesis	StALS1	Agrobacterium-mediated	U6	2×CaMV 35S	5–60	Butler et al. (2015)
	Knockout	Sb-CENH3	Agrobacterium-mediated	U6	ZmUbi	37–40	Che et al. (2018)
	Functional genomics	DsRED2	Agrobacterium-mediated	U6	OsActin1	NA	Jiang et al. (2013)
Rice (Oryza sativa)	Genome editing	TaLOX2	Agrobacterium-mediated	OsU3	2×CaMV 35S	∼1	Shan et al. (2014)
	Gene editing	OsPMS3, OsYSAOsDERF1, OsMYB1, OsMSH1, OsPDS, OsSPP, OsEPSPS, OsMYB5, and OsROC5	Agrobacterium-mediated	U3 and U6	ZmUbi. CaMV 35S	>35	Ma et al. (2015)
	Gene editing	SBEI and SBEIIb	Agrobacterium-mediated	OsU3	Ubi1	26.7–40	Sun et al. (2017)
	Knockout	elF4G	Agrobacterium-mediated	TaU6	ZmUbi1	30–64	Macovei et al. (2018)
	Knockout	OsNramp5	Agrobacterium-mediated	OsU6 and OsU3	Ubi1	13.6–35	Tang et al. (2017)
	Loss of function	OsMORE1 and OsMORE1a	Agrobacterium-mediated	OsU3	ZmUbi	NA	Kim et al. (2022)
	Knockout	SAPK1 and SAPK2	Agrobacterium-mediated	OsU3 and OsU6a	CaMV 35S	NA	Lou et al. (2018)
	Gene editing	OsPIN5b, gs3, and OsMYB30	Agrobacterium-mediated	U6	CaMV 35S	42–66	Zeng et al. (2020)
	Deletion	Waxya and Waxyb	Agrobacterium-mediated	OsU3 or OsU6	CaMV 35S	8.6–11.85	Liu et al. (2022)
	Gene editing	RLK	Agrobaterium-mediated	U3	Ubi1	NA	Chen et al. (2022)
	Knockout	ISA1	Agrobacterium-mediated	U6	CaMV 35S	NA	Shufen et al. (2019)
	Knockout	Waxy	Agrobacterium-mediated	U6	CaMV 35S	82.7–86.9	Zhang et al. (2018)
	Knockout	Waxy	Agrobacterium-mediated	U3 and U6a	CaMV 35S	NA	Yunyan et al. (2019)
	Knockout	OsRR22	Agrobacterium-mediated	OsU6a	UbiH	64.3	Zhang et al. (2019a)
	Knockout	OsCCD7	Agrobacterium-mediated	OsU3	OsUbi	22.2–64.3	Butt et al. (2018)
	Knockout	EPSPS	Protoplast transformation	OsU3 and TaU3	__	2.0–2.2	Li et al. (2016)
	Site-directed mutagenesis	OsROC5, OsSPP, and OsYSA	Agrobacterium-mediated	OsU6-2	CaMV 35S	61.1–67.7	Zhang et al. (2014)
	Site-directed mutagenesis	OsMPK5	Agrobacterium-mediated	U3 and U6	CaMV 35S	3–8	Xie and Yang. (2013)
	Site-directed mutagenesis	OsPDS, TaLOX2, OsBADH, and OsMPK2	Agrobacterium-mediated	OsU3	CaMV 35S	∼1	Shan et al. (2014)
	Site-directed mutagenesis	OsMYB1	Protoplast transformation	OsU3	CaMV 35S	NA	Miao et al. (2013)
	Multiplex editing capability with endogenous tRNA	OsMPKs	Agrobacterium-mediated	OsU3	OsUbi	6–100	Xie et al. (2015)
	Multiplex GE in dicot andmonocot plants	46 genomic targets	Agrobacterium-mediated	OsU3, OsU6 and OsU6c	OsUbi and CaMV 35S	24.7–90	Ma et al. (2015)
Barley (Hordeum vulgare)	Functional studies	WDV1, WDV2, WDV3, and WDV4	Agrobacterium-mediated	WDV	CaMV 35S and ZmUbi	NA	Kis et al. (2019)
	Fragment Deletions and Small Indels	ENGase	Agrobacterium-mediated	OsU6	ZmUbi	78	Kapusi et al. (2017)
	Gene editing	NbPDS1	Agrobacterium-mediated	U6	CaMV 35S	NA	Raitskin et al. (2019)
	Gene editing	Hpt	Agrobacterium- mediated	U6	ZmUbi	20–70	Watanabe et al. (2016)
	Knockout	HptII	Agrobacterium-mediated	U6	ZmUbi	NA	Lawrenson and Harwood. (2019)
	Knockout	HvMORC1 and HvMORC6a	Agrobacterium-mediated	HvU3	CaMV 35S	8–81	Galli et al. (2022)
	Knockout	HvCKX1	Agrobacterium-mediated	HvU3	ZmUbi1	NA	Holubova et al. (2018)
	Knockout	HvMORC1	Agrobacterium-mediated	HvU3	ZmUbi	38–77.7	Kumar et al. (2018)
	Knockout	HvCKX1, HvCKX3, and Nud	Agrobacterium-mediated	TaU6	ZmUbi	18–68	Gasparis et al. (2018)
	Knockout	GPhsp70, GPhsp26, GPhsp16.9, GPgst, GPcrt, GPipi, and GPpdi	Agrobacterium-mediated	TaU6	ZmUbi	NA	Panting et al. (2021)
Maize (Zea mays)	Knockout	waxy	Biolistic	TaU6	ZmUbi	1–55.4	Gao et al. (2020)
	Gene editing	ZmIPK	Agrobacterium-mediated	ZmU3	CaMV 35S	13.1	Liang et al. (2014)
	Gene editing	ZmHKT1	Agrobacterium-mediated	AtU6-26, OsU3 or TaU3	CaMV 35S or Ubi1	NA	Xing et al. (2014)
	Gene editing	LIG1, MS26, and MS45	Agrobacterium-mediated	ZmU6	ZmUbi	6–86	Svitashev et al. (2015)
	Gene editing	Zmzb7	Agrobacterium-mediated	ZmU3	CaMV 35S	19–31	Feng et al. (2016)
	Gene editing	PSY1	Agrobacterium-mediated	ZmU6	ZmUbi	10.67	Zhu et al. (2016)
	Gene editing	CLE	Agrobacterium-mediated	__	__	NA	Liu et al. (2021)