Efficient targeted mutagenesis by endogenous promoter controlled tRNA-gRNA array of CRISPR/Cas9 in potato and application in Solanum etuberosum

doi:10.21203/rs.3.rs-1665829/v1

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Efficient targeted mutagenesis by endogenous promoter controlled tRNA-gRNA array of CRISPR/Cas9 in potato and application in Solanum etuberosum

https://doi.org/10.21203/rs.3.rs-1665829/v1

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posted 07 Jun, 2022

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Here, we developed an endogenous promoter controlled polycistronic tRNA-gRNA (PTG) array of CRISPR/Cas9 in potato for efficient multiplex mutagenesis. The editing efficiency was significantly improved in potato combined with short-term heat treatment. In addition, our use of CRISPR/Cas9 to engineer mutations in wild solanum species demonstrates a high editing efficiency of the developed PTG/Cas9 in Solanum etuberosum, which contributes to develop Solanum etuberosum as a new model for functional genetics research in solanum.

CRISPR

gene editing

high temperature

potato

SP5G

Solanum etuberosum

Development of endogenous promoter driven CRISPR/Cas9 for high efficient gene editing in potato and Solanum etuberosum in combination with heat treatments.

Acknowledgements we are grateful to Kabin Xie from Huazhong agricultural university for the providing vector pRGEB32 and pGTR. This work was supported by the National Natural Science Foundation of China (No.31971988) and China Agriculture Research System (Potato, CARS-09-P08).

Author contribution statement S.J.,J.D. andB.S. conceived and designed research. S.J., W.X., H.L., F.D. conducted experiments. S.J., W.X., andB.S. analyzed the data. S.J. wrote the manuscript. K.C. and J.D. directed the study and revised the manuscript. All authors read and approved the manuscript.

Conflict of interest the authors declare no conflict of interest.

Abelenda JA, Cruz-Oro E, Franco-Zorrilla JM, Prat S (2016) Potato StCONSTANS-like1 Suppresses Storage Organ Formation by Directly Activating the FT-like StSP5G Repressor. Curr Biol 26:872–881
Gao CX (2021) Genome engineering for crop improvement and future agriculture. Cell 184:1621–1635
Guerineau F, Waugh R (1993) The U6 Small Nuclear-Rna Gene Family of Potato. Plant Mol Biol 22:807–818
LeBlanc C, Zhang F, Mendez J, Lozano Y, Chatpar K, Irish VF, Jacob Y (2018) Increased efficiency of targeted mutagenesis by CRISPR/Cas9 in plants using heat stress. Plant J 93:377–386
Li TD, Yang XP, Yu Y, Si XM, Zhai XW, Zhang HW, Dong WX, Gao CX, Xu C (2018) Domestication of wild tomato is accelerated by genome editing. Nat Biotechnol 36:1160–1163
Soyk S, Muller NA, Park SJ, Schmalenbach I, Jiang K, Hayama R, Zhang L, Van Eck J, Jimenez-Gomez JM, Lippman ZB (2017) Variation in the flowering gene SELF PRUNING 5G promotes day-neutrality and early yield in tomato. Nat Genet 49:162–168
Stokstad E (2019) The new potato. Science 363:574–577
Tang XM, Chen SL, Yu HW, Zheng XJ, Zhang F, Deng XX, Xu Q (2021) Development of a gRNA-tRNA array of CRISPR/Cas9 in combination with grafting technique to improve gene-editing efficiency of sweet orange. Plant Cell Rep 40:2453–2456
Xie KB, Minkenberg B, Yang YN (2015) Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. P Natl Acad Sci USA 112:3570–3575
Zhang SB, Jiao ZC, Liu L, Wang KT, Zhong DY, Li SB, Zhao TT, Xu XY, Cui X (2018) Enhancer-Promoter Interaction of SELF PRUNING 5G Shapes Photoperiod Adaptation. Plant Physiol 178:1631–1642

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Efficient targeted mutagenesis by endogenous promoter controlled tRNA-gRNA array of CRISPR/Cas9 in potato and application in Solanum etuberosum

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