An efficient CRISPR vector toolbox for engineering large deletions in Arabidopsis thaliana
| Autor: | Wangsheng Zhu, Joffrey Fitz, Efthymia Symeonidi, Detlef Weigel, Miriam Lucke, Rui Wu, Congmao Wang, Wanyan Xi, Yun-ting Jang, Rebecca Schwab |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Arabidopsis thaliana Plant Science Computational biology lcsh:Plant culture 01 natural sciences Genome 03 medical and health sciences NLR genes Genome editing Genetic variation Genetics CRISPR lcsh:SB1-1110 Vector (molecular biology) CRISPR/Cas9 lcsh:QH301-705.5 Gene biology Cas9 biology.organism_classification 030104 developmental biology lcsh:Biology (General) Natural variation 010606 plant biology & botany Biotechnology |
| Zdroj: | Plant Methods, Vol 14, Iss 1, Pp 1-9 (2018) |
| ISSN: | 1746-4811 |
| Popis: | Background Our knowledge of natural genetic variation is increasing at an extremely rapid pace, affording an opportunity to come to a much richer understanding of how effects of specific genes are dependent on the genetic background. To achieve a systematic understanding of such GxG interactions, it is desirable to develop genome editing tools that can be rapidly deployed across many different genetic varieties. Results We present an efficient CRISPR/Cas9 toolbox of super module (SM) vectors. These vectors are based on a previously described fluorescence protein marker expressed in seeds allowing identification of transgene-free mutants. We have used this vector series to delete genomic regions ranging from 1.7 to 13 kb in different natural accessions of the wild plant Arabidopsis thaliana. Based on results from 53 pairs of sgRNAs targeting individual nucleotide binding site leucine-rich repeat (NLR) genes, we provide a comprehensive overview of obtaining heritable deletions. Conclusions The SM series of CRISPR/Cas9 vectors enables the rapid generation of transgene-free, genome edited plants for a diversity of functional studies. |
| Databáze: | OpenAIRE |
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