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

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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Hong Yang, Jia-Jing Wu, Ting Tang, Ke-De Liu, Cheng Dai
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2db46f45aa1743b2b342601f7beb2209
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:2db46f45aa1743b2b342601f7beb2209
record_format	dspace
spelling	oai:doaj.org-article:2db46f45aa1743b2b342601f7beb22092021-12-02T15:05:15ZCRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus10.1038/s41598-017-07871-92045-2322https://doaj.org/article/2db46f45aa1743b2b342601f7beb22092017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-07871-9https://doaj.org/toc/2045-2322Abstract 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.Hong YangJia-Jing WuTing TangKe-De LiuCheng DaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hong Yang Jia-Jing Wu Ting Tang Ke-De Liu Cheng Dai CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus
description	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.
format	article
author	Hong Yang Jia-Jing Wu Ting Tang Ke-De Liu Cheng Dai
author_facet	Hong Yang Jia-Jing Wu Ting Tang Ke-De Liu Cheng Dai
author_sort	Hong Yang
title	CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus
title_short	CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus
title_full	CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus
title_fullStr	CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus
title_full_unstemmed	CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus
title_sort	crispr/cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgrna in brassica napus
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/2db46f45aa1743b2b342601f7beb2209
work_keys_str_mv	AT hongyang crisprcas9mediatedgenomeeditingefficientlycreatesspecificmutationsatmultiplelociusingonesgrnainbrassicanapus AT jiajingwu crisprcas9mediatedgenomeeditingefficientlycreatesspecificmutationsatmultiplelociusingonesgrnainbrassicanapus AT tingtang crisprcas9mediatedgenomeeditingefficientlycreatesspecificmutationsatmultiplelociusingonesgrnainbrassicanapus AT kedeliu crisprcas9mediatedgenomeeditingefficientlycreatesspecificmutationsatmultiplelociusingonesgrnainbrassicanapus AT chengdai crisprcas9mediatedgenomeeditingefficientlycreatesspecificmutationsatmultiplelociusingonesgrnainbrassicanapus
_version_	1718388881132355584

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

Ejemplares similares