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Cited by 199CRISPR and Genetic EngineeringPlant Genetic and Mutation StudiesGenetically Modified Organisms Research

CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus

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Yang, Hong, Wu, Jia-Jing, Tang, Ting, Liu, Ke-De, Dai, Cheng·2017·Scientific Reports
DOI 10.1038/s41598-017-07871-9PMC PMC5548805
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Abstract

CRISPR/Cas9 is a valuable tool for both basic and applied research that has been widely applied to different plant species. Nonetheless, a systematical assessment of the efficiency of this method is not available for the allotetraploid Brassica napus—an important oilseed crop. In this study, we examined the mutation efficiency of the CRISPR/Cas9 method for 12 genes and also determined the pattern, specificity and heritability of these gene modifications in B. napus. The average mutation frequency for a single-gene targeted sgRNA in the T0 generation is 65.3%. For paralogous genes located in conserved regions that were targeted by sgRNAs, we observed mutation frequencies that ranged from 27.6% to 96.6%. Homozygotes were readily found in T0 plants. A total of 48.2% of the gene mutations, including homozygotes, bi-alleles, and heterozygotes were stably inherited as classic Mendelian alleles in the next generation (T1) without any new mutations or reversions. Moreover, no mutation was found in the putative off-target sites among the examined T0 plants. Collectively, our results demonstrate that CRISPR/Cas9 is an efficient tool for creating targeted genome modifications at multiple loci that are stable and inheritable in B. napus. These findings open many doors for biotechnological applications in oilseed crops.

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Cited by

Sorted by citations

Authors
Yang, H., Wu, J.-J., Tang, T., Liu, K.-D., & Dai, C.
Affiliations
Huazhong Agricultural University (CN)
Corresponding author
Dai, Cheng
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2017-08-08
Volume
7
Issue
1
Pages
7489-7489
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/s41598-017-07871-9
PMC
PMC5548805
OpenAlex
W2742124987
Cited by
199
References
1
Citations by year
2026: 9 · 2025: 13 · 2024: 21 · 2023: 21 · 2022: 31 · 2021: 33 · 2020: 29 · 2019: 24 · 2018: 17 · 2017: 1
FWCI
7.93
Type
journal article
Language
en
Update notices
correction (2018-03-15)
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Natural Science Foundation of Hubei Province
  • Huazhong Agricultural University (0900206305)
  • National Key Research and Development Program of China (2016YFD0101007)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Genetic and Mutation Studies
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Genetically Modified Organisms Research
Keywords
CRISPR/Cas9, genome editing, Brassica napus, mutation efficiency, single guide RNA, paralogs, off-target mutagenesis, Mendelian inheritance, homozygous mutants, oilseed crops, allotetraploid
MeSH terms
Gene Editing, CRISPR-Associated Protein 9, RNA, Guide, CRISPR-Cas Systems, Alleles, Amino Acid Sequence, Base Sequence, Genotype, Homozygote, Phenotype, Sequence Alignment, Genes, Plant, Brassica napus, Genetic Loci, Tetraploidy, Mutation Rate, Agrobacterium, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems

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