The DOI matches its Crossref record; 8 of the 8 references it cites were found in a registry or an index.
The CRISPR/Cas system can be used as nuclease for in planta gene targeting and as paired nickases for directed mutagenesis in Arabidopsis resulting in heritable progeny
Abstract
Summary: The CRISPR/C as nuclease is becoming a major tool for targeted mutagenesis in eukaryotes by inducing double‐strand breaks (DSB s) at pre‐selected genomic sites that are repaired by non‐homologous end joining (NHEJ) in an error‐prone way. In plants, it could be demonstrated that the C as9 nuclease is able to induce heritable mutations in A rabidopsis thaliana and rice. Gene targeting (GT) by homologous recombination (HR) can also be induced by DSB s. Using a natural nuclease and marker genes, we previously developed an in planta GT strategy in which both a targeting vector and targeting locus are activated simultaneously via DSB induction during plant development. Here, we demonstrate that this strategy can be used for natural genes by CRISPR/C as‐mediated DSB induction. We were able to integrate a resistance cassette into the ADH 1 locus of A. thaliana via HR. Heritable events were identified using a PCR‐based genotyping approach, characterised by S outhern blotting and confirmed on the sequence level. A major concern is the specificity of the CRISPR/C as nucleases. Off‐target effects might be avoided using two adjacent sg RNA target sequences to guide the C as9 nickase to each of the two DNA strands, resulting in the formation of a DSB. By amplicon deep sequencing, we demonstrate that this C as9 paired nickase strategy has a mutagenic potential comparable with that of the nuclease, while the resulting mutations are mostly deletions. We also demonstrate the stable inheritance of such mutations in A. thaliana.
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Cites
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J. A., & Charpentier, E. (2012) · Science · doi:10.1126/science.1225829 · Cited by 17,116
High-Frequency off-Target Mutagenesis Induced by CRISPR-Cas Nucleases in Human Cells
Fu, Y., Foden, J. A., Khayter, C., Maeder, M. L., Reyon, D., Joung, J. K., & Sander, J. D. (2013) · Nature Biotechnology · doi:10.1038/nbt.2623 · Cited by 3,292
RNA-guided genetic silencing systems in bacteria and archaea
Wiedenheft, B., Sternberg, S. H., & Doudna, J. A. (2012) · Nature · doi:10.1038/nature10886 · Cited by 1,851
Mali, P., Aach, J., Stranges, P. B., Esvelt, K. M., Moosburner, M., Kosuri, S., Yang, L., & Church, G. M. (2013) · Nature Biotechnology · doi:10.1038/nbt.2675 · Cited by 1,752
Veres, A., Gosis, B. S., Ding, Q., Collins, R., Ragavendran, A., Brand, H., Erdin, S., Cowan, C. A., Talkowski, M. E., & Musunuru, K. (2014) · Cell Stem Cell · doi:10.1016/j.stem.2014.04.020 · Cited by 474
Precision genome engineering and agriculture: opportunities and regulatory challenges
Voytas, D. F., & Gao, C. (2014) · PLoS Biology · doi:10.1371/journal.pbio.1001877 · Cited by 458
Smith, C., Gore, A., Yan, W., Abalde-Atristain, L., Li, Z., He, C., Wang, Y., Brodsky, R. A., Zhang, K., Cheng, L., et al. (2014) · Cell stem cell · doi:10.1016/j.stem.2014.06.011 · Cited by 342
Hartung, F., & Schiemann, J. (2014) · The Plant Journal · doi:10.1111/tpj.12413 · Cited by 277
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A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation
Lowder, L. G., Zhang, D., Baltes, N. J., Paul, J. W., Tang, X., Zheng, X., Voytas, D. F., Hsieh, T.-F., Zhang, Y., & Qi, Y. (2015) · Plant Physiology · doi:10.1104/pp.15.00636 · Cited by 707
Editing plant genomes with CRISPR/Cas9
Belhaj, K., Chaparro-Garcia, A., Kamoun, S., Patron, N. J., & Nekrasov, V. (2015) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2014.11.007 · Cited by 537
Genetically modified crops: current status and future prospects
Kumar, K., Gambhir, G., Dass, A., Tripathi, A. K., Singh, A., Jha, A. K., Yadava, P., Choudhary, M., & Rakshit, S. (2020) · Planta · doi:10.1007/s00425-020-03372-8 · Cited by 528
SCFTIR1/AFB-Based Auxin Perception: Mechanism and Role in Plant Growth and Development
Salehin, M., Bagchi, R., & Estelle, M. (2015) · The Plant Cell · doi:10.1105/tpc.114.133744 · Cited by 462
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Metadata
- Authors
- Schiml, S., Fauser, F., & Puchta, H.
- Venue
- The Plant Journal
- Publisher
- Wiley
- Published
- 2014-11-11
- Volume
- 80
- Issue
- 6
- Pages
- 1139-1150
- ISSN
- 0960-7412 (ISSN-L)
- 1365-313X
- OpenAlex
- W2155194596
- Cited by
- 342
- References
- 8
- FWCI
- 14.84
- Type
- journal article
- Language
- en
- Open access
- closed
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- Schiml, S., Fauser, F., & Puchta, H. (2014). The CRISPR/Cas system can be used as nuclease for in planta gene targeting and as paired nickases for directed mutagenesis in Arabidopsis resulting in heritable progeny. The Plant Journal. https://doi.org/10.1111/tpj.12704
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