Genome Editing in Plants: An Overview of Tools and Applications

Table 1

Comparison of plant genome editing techniques.

	ZFNs	TALENs	ODM	CRISPR/Cas9	Reference	Year
Components	Zn finger domains; Nonspecific FokI nuclease domain	TALE DNA-binding domains; Nonspecific FokI nuclease domain	Exogenous polynucleotide (chimeraplast)	crRNA, Cas9 proteins	Kumar and Jain [6]; Sauer et al. [7]	2015
						2016
Structural proteins	Dimeric protein	Dimeric protein	Nonprotein nature	Monomeric protein	Li et al. [8]	2013
					Gaj et al. [4]	2013
					Sauer et al. [7]	2016
Catalytic domain	Restriction endonuclease FokI	Restriction endonuclease FokI	There is no catalytic domain	RUVC and HNH	Jinek et al. [9]	2012
					Sauer et al. [7]	2016
Length of target sequence (bp)	24–36	24–59	68–88	20–22	Gaj et al. [4]	2013
					Chen et al. [10]	2016
					Sauer et al. [7]	2016
Protein engineering steps	Required	Required	Not required	Should not be complex to test gRNA	Cho et al. [11]	2013
					Weeks et al. [5]	2016
					Sauer et al. [7]	2016
Cloning	Necessary	Necessary	Not necessary	Not necessary	Cho et al. [11]	2013
					Weeks et al. [5]	2016
					Sauer et al. [7]	2016
gRNA production	Not applicable	Not applicable	Not required	Easy to produce	Cho et al. [11]	2013
					Noman et al. [12]	2016
					Sauer et al. [7]	2016
Mode of action	Double-strand breaks in target DNA	Double-strand breaks in target DNA	Information strand directs conversion(s) within target region	Double-strand breaks or single-strand nicks in target DNA	Li et al. [8]	2013
					Mao et al. [13]	2013
					Noman et al. [12]	2016
					Sauer et al. [7]	2016
Target recognition efficiency	High	High	High	High	Gaj et al. [4]	2013
					Kumar and Jain [6]	2015
					Sauer et al. [7]	2016
Mutation rate	High	Middle	Middle	Low	Li et al. [8]	2013
					Gaj et al. [4]	2013
					Sauer et al. [7]	2016
Creation of large scale libraries	Impossible	Technically difficult	Technically difficult	Possible	Hsu [14]	2013
					Cho et al. [11]	2013
					Sauer et al. [7]	2016
Multiplexing	Difficult	Difficult	Technically difficult	Possible	Li et al. [8]	2013
					Mao et al. [13]	2013
					Noman et al. [12]	2016
					Sauer et al. [7]	2016