Table 1 Methods for the establishment of the null segregants
From: Precision genome editing in plants: state-of-the-art in CRISPR/Cas9-based genome engineering
Methods | Plant materials | Transformation methods | Cas9 and gRNA delivery | Reference |
|---|---|---|---|---|
Mendelian segregation (supporeted by new screening strategies) | Arabidopsis | Agrobacterium-mediated transformation (floral dipping) | Encoding DNA | [35] |
Rice calli | Agrobacterium-mediated transformation | Encoding DNA | [34] | |
Programmed self-elimination of transgenic plants | Rice calli | Agrobacterium-mediated transformation | Encoding DNA | [36] |
Transient expression of CRISPR/Cas9 from DNA or mRNA | Potato protoplasts | PEG-mediated transformation | Encoding DNA | [37] |
B. oldhamii, S. italica, O. sativa, Z. mays, A. thaliana, B. oleracea, B. napus, N. tabacum, S. lycopersicum protoplasts | PEG-mediated transformation | Encoding DNA | [38] | |
Tobacco leaf explants | Agrobacterium-mediated transformation | Encoding DNA | [39] | |
Wheat calli | Particle bombardment | Encoding DNA or RNA | [40] | |
RNPs-mediated targeted mutagenesis | Arabidopsis, tobacco, lettuce and rice protoplasts | PEG-mediated transformation | Protein and in vitro-transcribed gRNA | [41] |
Grape, apple protoplasts | PEG-mediated transformation | Protein and in vitro-transcribed gRNA | [42] | |
Petunia × hybrida protolasts | PEG-mediated transformation | Protein and in vitro-transcribed gRNA | [43] | |
Wheat protoplast, immmature embryo | PEG-mediated transformation, particle bombardment | Protein and in vitro-transcribed gRNA | [44] | |
B. oleracea and B. rapa protoplasts | PEG-mediated transformation | Protein and in vitro-transcribed gRNA or synthesized gRNA | [45] | |
Maize immmature embryo | Particle bombardment | Protein and in vitro-transcribed gRNA | [46] | |
Rice zygote produced by in vitro ferlization of isolated gamates | PEG-mediated transformation | Protein and in vitro-transcribed gRNA | [47] |