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Table 1 Summary of CRISPR/Cas genome editing assays in plants

From: Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system

Material/activity A. thaliana N. benthamiana O. sativa T. aestivum S. bicolor Reference
Cas9
Codon-optimized for plants Yes Yes Yes Yes Yes [10, 11, 15, 16]
No No No    [9, 1214, 16]
Number of NLS 2 2 2 2   [1014]
  1     [9, 16]
1   1   N/A [16]
   1    [7, 8, 15]
Intron introduced Yes Yes     [11]
No No No No No [9, 10, 1216]
Promoter 35S PPDK 35S PPDK     [11]
  2x CaMV 35S   2x CaMV 35S 2x CaMV 35S   [10, 12]
  CaMV 35S CaMV 35S CaMV 35S    [9, 11, 14, 16]
      OsAct1 [16]
  AtUBQ   OsUBQ    [13]
    ZmUBQ    [15]
sgRNA
Promoter AtU6 AtU6     [9, 1113, 16]
    OsU3    [10, 1215]
    OsU6   OsU6 [16]
     TaU6   [10]
Assays
  AtPDS3   OsPDS    
  AtFLS2   OsBADH2    
  AtBRI1   OsMPK2    
  AtJAZ1   Os02g23823    
  AtGAI   OsROC5    
  AtCHL1   OsSPP    
Genes targeted AtCHL2 NbPDS OsYSA TaMLO N/A [916]
  At5g13930   OsMYB1    
    OsMPK5    
    OsCAO1    
    OsLAZY1    
    OsSWEET11    
    OsSWEET14    
Transient assays
Protoplasts Yes Yes Yes Yes No [1014, 16]
NHEJ mutation frequency 1.1-5.6% [11] 37.7-38.5% [11] 14.5-38% [10] 28-29% [10]   
    3-8% [14]    
HDR modification frequency 18.8% [12]* 9% [11] 6.9% [10]    
  42% [13]*      
Leaf agroinfiltration Yes Yes No No No [9, 11, 16]
NHEJ mutation frequency 2.7% [11] 4.8% [11]     
   2.1% [9]     
Embryo transformation No No No No Yes [16]
NHEJ mutation frequency      28%  
Transgenic mutants
Mutated plants recovered Yes Yes Yes N/A N/A [9, 10, 12, 13, 15]
Frequency of modified plants 30-84% [12] 6.7% [9] 4-9.4% [10]    
  76-89% [13]   5-75% [12]    
    50% [13]    
    83-91.6% [15]    
Biallelic mutations recovered Yes [12, 13] No [9, 11] Yes [10, 12, 14, 15] N/A N/A  
Off-targets
Off-target detected N/A No [9] Yes [10, 14] N/A N/A [9, 10, 14]
  1. *In YF-FP assays.
  2. Abbreviations: A. thaliana: Arabidopsis thaliana, N. benthamiana: Nicotiana benthamiana, O. sativa: Oryza sativa, T. aestivum: Triticum aestivum, S. bicolor: Sorghum bicolor.