RECOMMENDED READINGCRISPR-Cas9 gene
editing in plants

The following are a list of publications that demonstrate the applicationof CRISPR-Cas9 genome editing techniques for targeted gene knockout orprecise knockin in a variety of plant species.
1. Svitashev, S., C. Schwartz, et al. Genome editing in maize directed by
CRISPR–Cas9 ribonucleoprotein complexes. Nat. Commun. 7, (2016).
[Zea mays – particle bombardment of embryos]
2. Svitashev, S., J. Young, et al. Targeted Mutagenesis, Precise Gene Editing,and Site-Specific Gene Insertion in Maize Using Cas9 and Guide RNA. Plant
Physiol. 2, 931-945 (2015). [Zea mays – particle bombardment of embryos]
3. Woo, J.W., J. Kim, et al. DNA-free genome editing in plants withpreassembled CRISPR-Cas9 ribonucleoproteins. Nat. Biotechnol. 33, 11621164 (2015). [Arabidopsis, Oryza sativa, and Lactuca sativa – protoplasttransfection]
4. Subburaj, S., S. J. Chung, et al. Site-directed mutagenesis in Petunia ×
hybrida protoplast system using direct delivery of purified recombinantCas9 ribonucleoproteins. Plant Cell Rep. 7, 1535-1544 (2016). [Petunia ×hybrida – protoplast transfection]
5. Zhang, Y., Z. Liang, et al. Efficient and transgene-free genome editing inwheat through transient expression of CRISPR/Cas9 DNA or RNA.
Nat. Commun. 7, (2016). [Triticum aestivum – particle bombardment ofprotoplasts]
6. Malnoy, M., R. Viola, et al. DNA-Free Genetically Edited Grapevine andApple Protoplast Using CRISPR/Cas9 Ribonucleoproteins. Front. Plant Sci.
7, (2016). [Chardonnay and Golden Delicious – protoplast transformation]
7. Baek, K. D. H. Kim, et al. DNA-free two-gene knockout in Chlamydomonas
reinhardtii via CRISPR-Cas9 ribonucleoproteins. Sci. Rep. 6, (2016).
[Chlamydomonas reinhardtii – transformation]
8. Li, J.F., J.E. Norville, et al. Multiplex and homologous recombinationmediated genome editing in Arabidopsis and Nicotiana benthamianausing guide RNA and Cas9. Nat. Biotechnol. 31, 688-691 (2013).
[Arabidopsis thaliana and Nicotiana benthamiana - PEG-protoplasttransfection, leaf agroinfiltration]
9. Jiang, W., H. Zhou, et al. Demonstration of CRISPR/Cas9/sgRNA-mediatedtargeted gene modification in Arabidopsis, tobacco, sorghum andrice. Nucleic Acids Res. 41, e188 (2013). [Arabidopsis thaliana, Nicotiana
benthamiana, Oryza sativa and Sorghum bicolor - leaf agroinfiltration, PEGprotoplast transfection]

10. Mao, Y., H. Zhang, et al. Application of the CRISPR-Cas System for
Efficient Genome Engineering in Plants. Mol. Plant. 6, 2008-2011 (2013).
[Arabidopsis thaliana and Oryza sativa - agro-transformation by floral
dip, stable agro-transformation]
11. Fauser, F., S. Schiml, et al. Both CRISPR/Cas-based nucleases and nickases
can be used efficiently for genome engineering in Arabidopsis thaliana.
Plant J. 79, 348-359 (2014). [Arabidopsis thaliana - agro-transformation byfloral dip, stable agro-transformation]
12. Peterson, B.A., D.C. Haak, et al. Genome-Wide Assessment of Efficiency
and Specificity in CRISPR/Cas9 Mediated Multiple Site Targeting in
Arabidopsis. PLoS One. 11, e0162169 (2016). [Arabidopsis thaliana – agrotransformation by floral dip]
13. Pyott, D.E., E. Sheehan, et al. Engineering of CRISPR/Cas9-mediatedpotyvirus resistance in transgene-free Arabidopsis plants. Mol. Plant
Pathol. 17, 1276-1288 (2016). [Arabidopsis thaliana – agro-transformation
by floral dip]
14. Nekrasov, V., B. Staskawicz, et al. Targeted mutagenesis in the model
plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease.
Nat. Biotechnol. 31, 691-693 (2013). [Nicotiana benthamiana - leaf
agroinfiltration]
15. Upadhyay, S.K., J. Kumar, et al. RNA-Guided Genome Editing for Target
Gene Mutations in Wheat. G3 (Bethesda) 3, 2233-2238 (2013). [Nicotianabenthamiana and Triticum aestivum - leaf agroinfiltration]
16. Shan, Q., Y. Wang, et al. Targeted genome modification of crop plants
using a CRISPR-Cas system. Nat. Biotechnol. 31, 686-688 (2013). [Oryzasativa and Triticum aestivum - PEG-protoplast transfection, particle
bombardment of callus]
17. Xie, K. and Y. Yang. RNA-guided genome editing in plants using aCRISPR-Cas system. Mol. Plant. 6, 1975-1983 (2013). [Oryza sativa – PEGprotoplast transfection]
18. Zhou, H. and B. Liu. Large chromosomal deletions and heritable small
genetic changes induced by CRISPR/Cas9 in rice. Nucleic Acids Res. 42,10903-10914 (2014). [Oryza sativa – PEG-protoplast transfection]
19. Miao, J., D. Guo, et al. Targeted mutagenesis in rice using CRISPR-Cas
system. Cell Res. 23, 1233-1236 (2013). [Oryza sativa – agro-transformationof callus, transient particle bombardment of callus]

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20. Baysal, C., L. Bortesi, et al. CRISPR/Cas9 activity in the rice OsBEIIb gene does
not induce off-target effects in the closely related paralog OsBEIIa. Mol.
Breeding. 36, (2016). [Oryza sativa - particle bombardment of embryos]
21. Brooks, C., V. Nekrasov, et al. Efficient Gene Editing in Tomato in the First
Generation Using the Clustered Regularly Interspaced Short Palindromic
Repeats/CRISPR-Associated9 System. Plant Physiol. 166, 1292-1297 (2014).
[Solanum lycopersicum - agro-transformation of cotyledons]
22. Ron, M., K. Kajala, et al. Hairy root transformation using Agrobacterium
rhizogenes as a tool for exploring cell type-specific gene expression andfunction using tomato as a model. Plant Physiol. 166, 455-469 (2014).
[Solanum lycopersicum - hairy root transformation by A. rhizogenes]
23. Pan, C., L. Ye, et al. CRISPR/Cas9-mediated efficient and heritable targetedmutagenesis in tomato plants in the first and later generations. Sci. Rep. 6(2016). [Solanum lycopersicum - agro-transformation of leaf discs]
24. Jia, H. and N. Wang. Targeted genome editing of sweet orange using Cas9/
sgRNA. PLoS One. 9, e93806 (2014). [Citrus sinensis - leaf agroinfiltration]
25. Sugano, S.S., M. Shirakawa, et al. CRISPR/Cas9-mediated targetedmutagenesis in the liverwort Marchantia polymorpha L. PlantCell Physiol. 55, 475-481 (2014). [Marchantia polymorpha - agrotransformation of sporelings]

26. Liang, Z., K. Zhang, et al. Targeted mutagenesis in Zea mays using TALENs
and the CRISPR/Cas system. J. Genet. Genomics. 41, 63-68 (2014).
[Zea mays – PEG-protoplast transfection]
27. Shi, J., H. Gao, et al. ARGOS8 variants generated by CRISPR-Cas9 improvemaize grain yield under field drought stress conditions. Plant Biotechnol. J.
(2016). [Zea mays – particle bombardment of embryos]
28. Nishitani, C., N. Hirai, et al. Efficient Genome Editing in Apple Usinga CRISPR/Cas9 system. Sci. Rep. 6, (2016). [Malus prunifolia – agrotransformation of shoots]
29. Fan, D., T. Liu, et al. Efficient CRISPR/Cas9-mediated Targeted Mutagenesis
in Populus in the First Generation. Sci. Rep. 5, (2015). [Populus tomentosa –agro-transformation of leaf discs]
30. Lawrenson, T., O. Shorinola, et al. Induction of targeted, heritable
mutations in barley and Brassica oleracea using RNA-guided Cas9nuclease. Genome Biol. 16, (2015). [Hordeum vulgare and Brassica
oleracea – agro-transformation of embryos, agro-transformation ofcotyledonary petioles]

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