Wuschel2 enables highly efficient CRISPR/Cas-targeted genome editing during rapid de novo shoot regeneration in sorghum
For many important crops including sorghum, use of CRISPR/Cas technology is limited not only by the delivery of the gene-modification components into a plant cell, but also by the ability to regenerate a fertile plant from the engineered cell through tissue culture. Here, we report that Wuschel2 (Wu...
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Published in | Communications biology Vol. 5; no. 1; p. 344 |
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Main Authors | , , , , , , , , , , |
Format | Journal Article |
Language | English |
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London
Nature Publishing Group UK
11.04.2022
Nature Publishing Group Nature Portfolio |
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Abstract | For many important crops including sorghum, use of CRISPR/Cas technology is limited not only by the delivery of the gene-modification components into a plant cell, but also by the ability to regenerate a fertile plant from the engineered cell through tissue culture. Here, we report that
Wuschel2 (Wus2)
-enabled transformation increases not only the transformation efficiency, but also the CRISPR/Cas-targeted genome editing frequency in sorghum (
Sorghum bicolor L
.). Using
Agrobacterium
-mediated transformation, we have demonstrated
Wus2
-induced direct somatic embryo formation and regeneration, bypassing genotype-dependent callus formation and significantly shortening the tissue culture cycle time. This method also increased the regeneration capacity that resulted in higher transformation efficiency across different sorghum varieties. Subsequently, advanced excision systems and “altruistic” transformation technology have been developed to generate high-quality morphogenic gene-free and/or selectable marker-free sorghum events. Finally, we demonstrate up to 6.8-fold increase in CRISPR/Cas9-mediated gene dropout frequency using
Wus2
-enabled transformation, compared to without
Wus2
, across various targeted loci in different sorghum genotypes.
Che et al. use Wuschel2-enabled genome transformation to induce somatic embryo formation in sorghum, a grain used in human food. Their approach not only overcomes the genotype-dependent barrier for genetic transformation without the introduction of morphogenic genes, but also increases the frequency of CRISPR/Castargeted genome editing. |
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AbstractList | For many important crops including sorghum, use of CRISPR/Cas technology is limited not only by the delivery of the gene-modification components into a plant cell, but also by the ability to regenerate a fertile plant from the engineered cell through tissue culture. Here, we report that Wuschel2 (Wus2)-enabled transformation increases not only the transformation efficiency, but also the CRISPR/Cas-targeted genome editing frequency in sorghum (Sorghum bicolor L.). Using Agrobacterium-mediated transformation, we have demonstrated Wus2-induced direct somatic embryo formation and regeneration, bypassing genotype-dependent callus formation and significantly shortening the tissue culture cycle time. This method also increased the regeneration capacity that resulted in higher transformation efficiency across different sorghum varieties. Subsequently, advanced excision systems and “altruistic” transformation technology have been developed to generate high-quality morphogenic gene-free and/or selectable marker-free sorghum events. Finally, we demonstrate up to 6.8-fold increase in CRISPR/Cas9-mediated gene dropout frequency using Wus2-enabled transformation, compared to without Wus2, across various targeted loci in different sorghum genotypes.Che et al. use Wuschel2-enabled genome transformation to induce somatic embryo formation in sorghum, a grain used in human food. Their approach not only overcomes the genotype-dependent barrier for genetic transformation without the introduction of morphogenic genes, but also increases the frequency of CRISPR/Castargeted genome editing. For many important crops including sorghum, use of CRISPR/Cas technology is limited not only by the delivery of the gene-modification components into a plant cell, but also by the ability to regenerate a fertile plant from the engineered cell through tissue culture. Here, we report that Wuschel2 (Wus2) -enabled transformation increases not only the transformation efficiency, but also the CRISPR/Cas-targeted genome editing frequency in sorghum ( Sorghum bicolor L .). Using Agrobacterium -mediated transformation, we have demonstrated Wus2 -induced direct somatic embryo formation and regeneration, bypassing genotype-dependent callus formation and significantly shortening the tissue culture cycle time. This method also increased the regeneration capacity that resulted in higher transformation efficiency across different sorghum varieties. Subsequently, advanced excision systems and “altruistic” transformation technology have been developed to generate high-quality morphogenic gene-free and/or selectable marker-free sorghum events. Finally, we demonstrate up to 6.8-fold increase in CRISPR/Cas9-mediated gene dropout frequency using Wus2 -enabled transformation, compared to without Wus2 , across various targeted loci in different sorghum genotypes. Che et al. use Wuschel2-enabled genome transformation to induce somatic embryo formation in sorghum, a grain used in human food. Their approach not only overcomes the genotype-dependent barrier for genetic transformation without the introduction of morphogenic genes, but also increases the frequency of CRISPR/Castargeted genome editing. Che et al. use Wuschel2-enabled genome transformation to induce somatic embryo formation in sorghum, a grain used in human food. Their approach not only overcomes the genotype-dependent barrier for genetic transformation without the introduction of morphogenic genes, but also increases the frequency of CRISPR/Castargeted genome editing. For many important crops including sorghum, use of CRISPR/Cas technology is limited not only by the delivery of the gene-modification components into a plant cell, but also by the ability to regenerate a fertile plant from the engineered cell through tissue culture. Here, we report that Wuschel2 (Wus2)-enabled transformation increases not only the transformation efficiency, but also the CRISPR/Cas-targeted genome editing frequency in sorghum (Sorghum bicolor L.). Using Agrobacterium-mediated transformation, we have demonstrated Wus2-induced direct somatic embryo formation and regeneration, bypassing genotype-dependent callus formation and significantly shortening the tissue culture cycle time. This method also increased the regeneration capacity that resulted in higher transformation efficiency across different sorghum varieties. Subsequently, advanced excision systems and "altruistic" transformation technology have been developed to generate high-quality morphogenic gene-free and/or selectable marker-free sorghum events. Finally, we demonstrate up to 6.8-fold increase in CRISPR/Cas9-mediated gene dropout frequency using Wus2-enabled transformation, compared to without Wus2, across various targeted loci in different sorghum genotypes. Abstract For many important crops including sorghum, use of CRISPR/Cas technology is limited not only by the delivery of the gene-modification components into a plant cell, but also by the ability to regenerate a fertile plant from the engineered cell through tissue culture. Here, we report that Wuschel2 (Wus2) -enabled transformation increases not only the transformation efficiency, but also the CRISPR/Cas-targeted genome editing frequency in sorghum ( Sorghum bicolor L .). Using Agrobacterium -mediated transformation, we have demonstrated Wus2 -induced direct somatic embryo formation and regeneration, bypassing genotype-dependent callus formation and significantly shortening the tissue culture cycle time. This method also increased the regeneration capacity that resulted in higher transformation efficiency across different sorghum varieties. Subsequently, advanced excision systems and “altruistic” transformation technology have been developed to generate high-quality morphogenic gene-free and/or selectable marker-free sorghum events. Finally, we demonstrate up to 6.8-fold increase in CRISPR/Cas9-mediated gene dropout frequency using Wus2 -enabled transformation, compared to without Wus2 , across various targeted loci in different sorghum genotypes. |
ArticleNumber | 344 |
Author | Che, Ping Albertsen, Marc C. Gordon-Kamm, William Anand, Ajith Simon, Marissa K. Jones, Todd J. Yang, Meizhu Wu, Emily Gao, Huirong Sigmund, Amy L. Lowe, Keith |
Author_xml | – sequence: 1 givenname: Ping orcidid: 0000-0001-6074-2266 surname: Che fullname: Che, Ping email: ping.che@corteva.com organization: Corteva Agriscience – sequence: 2 givenname: Emily surname: Wu fullname: Wu, Emily organization: Corteva Agriscience – sequence: 3 givenname: Marissa K. surname: Simon fullname: Simon, Marissa K. organization: Corteva Agriscience – sequence: 4 givenname: Ajith surname: Anand fullname: Anand, Ajith organization: Corteva Agriscience – sequence: 5 givenname: Keith surname: Lowe fullname: Lowe, Keith organization: Corteva Agriscience – sequence: 6 givenname: Huirong orcidid: 0000-0001-8654-7661 surname: Gao fullname: Gao, Huirong organization: Corteva Agriscience – sequence: 7 givenname: Amy L. surname: Sigmund fullname: Sigmund, Amy L. organization: Corteva Agriscience – sequence: 8 givenname: Meizhu surname: Yang fullname: Yang, Meizhu organization: Corteva Agriscience – sequence: 9 givenname: Marc C. orcidid: 0000-0003-3058-2366 surname: Albertsen fullname: Albertsen, Marc C. organization: Corteva Agriscience – sequence: 10 givenname: William surname: Gordon-Kamm fullname: Gordon-Kamm, William organization: Corteva Agriscience – sequence: 11 givenname: Todd J. orcidid: 0000-0001-5452-5601 surname: Jones fullname: Jones, Todd J. organization: Corteva Agriscience |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/35410430$$D View this record in MEDLINE/PubMed |
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Snippet | For many important crops including sorghum, use of CRISPR/Cas technology is limited not only by the delivery of the gene-modification components into a plant... Abstract For many important crops including sorghum, use of CRISPR/Cas technology is limited not only by the delivery of the gene-modification components into... Che et al. use Wuschel2-enabled genome transformation to induce somatic embryo formation in sorghum, a grain used in human food. Their approach not only... |
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SubjectTerms | 13 13/106 42 42/41 631/449 631/61 Agrobacterium Biology Biomedical and Life Sciences Callus Cell culture CRISPR CRISPR-Cas Systems Edible Grain - genetics Gene Editing - methods Genetic transformation Genomes Genotypes Life Sciences Plants, Genetically Modified - genetics Regeneration - genetics Sorghum Sorghum - genetics Sorghum bicolor Tissue culture |
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Title | Wuschel2 enables highly efficient CRISPR/Cas-targeted genome editing during rapid de novo shoot regeneration in sorghum |
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