Table 4.

Different plasmids with specific Cas9 activity.

Crop	Plasmid	Gene/insert name	Promoter		Selectable Marker	Cas9 type	Significance	Reference
			sgRNA expression	Cas9 expression
Oryza sativa	pRGEB32	PTG1/Cas9	U3 snoRNA	Rice ubiquitin	Hygromycin	Cut	Enhanced multiplex editing capability via endogenous tRNA processing system	Xie et al., 2015
Zea mays	pHSE401	gRNA scaffold	AtU6-26	35S	Hygromycin	Cut	Improved designing of CRISPR/Cas9 binary vector. Easy method for one or more gRNA assembly in expression cassette, high efficiency mutant generation	Xing et al., 2014
Arabidopsis thaliana	pKI 1.1R	Human codon optimized spCas9	U6-26p	CaMV35S, WOX2, RPS5A	Hygromycin	Cut	pKIR vector harboring RPS5A maintains high constitutive expression and heritable muatations at all developmental stages	Tsutsui and Higashiyama, 2017
Arabidopsis, Nicotiana benthamiana, Oryza sativa	pYPQ159	hSpCas9D10A (human codon optimized)	AtU6-26	2× 35S	Spectinomycin	Nick	This toolbox provides reagents to efficiently assemble DNA constructs for monocots and dicots using Golden Gate cloning	Lowder et al., 2015
Zea mays, Arabidopsis	pBUN501	zCas9D10A	AtU6-26	Ubi1	Bar	Nick	Multiplex genome editing	Xing et al., 2014
Zea mays, Arabidopsis	pHSN6A01	dCas9-VP64, gRNA scaffold	AtU6-26p	2× 35Sp	Hygromycin	Activate	Multiplex genome editing	Xing et al., 2014
Zea mays, Arabidopsis	pBUN6A11	dCas9-VP64, gRNA scaffold	OsU3p	Ubi1p	Bar	Activate	Multiplex genome editing	Xing et al., 2014
Arabidopsis, Nicotiana benthamiana	pEGB 35S:dCas9:Tnos (GB1191)	dCas9	AtU6-26	35S, Tnos	Kanamycin	Interfere	GoldenBraid 2.0 validation- foster DNA exchange for synthetic biology; protein-protein interaction tool, gene silencing tool	Perdigones et al., 2013
Nicotiana benthamiana	pdCas9 (GB1079)	Mutated Cas9 (D10A, H840A) and inactivated catalytic domains (human codon optimized)	U6-26	pNOS	Ampicillin	Interfere	GoldenBraid gRNA-Cas9 toolbox validated; multiple TDNA cloning, gRNA multiplexing; effective transcriptional activation and repression	Vazquez-Vilar et al., 2016