High-efficiency CRISPR/Cas9 multiplex gene editing using the glycine tRNA-processing system-based strategy in maize
CRISPR/Cas9 genome editing strategy has been applied to a variety of species and the tRNA-processing system has been used to compact multiple gRNAs into one synthetic gene for manipulating multiple genes in rice. We optimized and introduced the multiplex gene editing strategy based on the tRNA-proce...
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| Veröffentlicht in: | BMC biotechnology 2016-08, Vol.16 (1), p.58-58, Article 58 |
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| creator | Qi, Weiwei Zhu, Tong Tian, Zhongrui Li, Chaobin Zhang, Wei Song, Rentao |
| description | CRISPR/Cas9 genome editing strategy has been applied to a variety of species and the tRNA-processing system has been used to compact multiple gRNAs into one synthetic gene for manipulating multiple genes in rice.
We optimized and introduced the multiplex gene editing strategy based on the tRNA-processing system into maize. Maize glycine-tRNA was selected to design multiple tRNA-gRNA units for the simultaneous production of numerous gRNAs under the control of one maize U6 promoter. We designed three gRNAs for simplex editing and three multiple tRNA-gRNA units for multiplex editing. The results indicate that this system not only increased the number of targeted sites but also enhanced mutagenesis efficiency in maize. Additionally, we propose an advanced sequence selection of gRNA spacers for relatively more efficient and accurate chromosomal fragment deletion, which is important for complete abolishment of gene function especially long non-coding RNAs (lncRNAs). Our results also indicated that up to four tRNA-gRNA units in one expression cassette design can still work in maize.
The examples reported here demonstrate the utility of the tRNA-processing system-based strategy as an efficient multiplex genome editing tool to enhance maize genetic research and breeding. |
| doi_str_mv | 10.1186/s12896-016-0289-2 |
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We optimized and introduced the multiplex gene editing strategy based on the tRNA-processing system into maize. Maize glycine-tRNA was selected to design multiple tRNA-gRNA units for the simultaneous production of numerous gRNAs under the control of one maize U6 promoter. We designed three gRNAs for simplex editing and three multiple tRNA-gRNA units for multiplex editing. The results indicate that this system not only increased the number of targeted sites but also enhanced mutagenesis efficiency in maize. Additionally, we propose an advanced sequence selection of gRNA spacers for relatively more efficient and accurate chromosomal fragment deletion, which is important for complete abolishment of gene function especially long non-coding RNAs (lncRNAs). Our results also indicated that up to four tRNA-gRNA units in one expression cassette design can still work in maize.
The examples reported here demonstrate the utility of the tRNA-processing system-based strategy as an efficient multiplex genome editing tool to enhance maize genetic research and breeding.</description><identifier>ISSN: 1472-6750</identifier><identifier>EISSN: 1472-6750</identifier><identifier>DOI: 10.1186/s12896-016-0289-2</identifier><identifier>PMID: 27515683</identifier><language>eng</language><publisher>England: BioMed Central Ltd</publisher><subject>breeding ; Clustered Regularly Interspaced Short Palindromic Repeats ; Corn ; CRISPR-Associated Proteins ; gene editing ; Gene Editing - methods ; Gene mutations ; Genes ; Genes, Plant - genetics ; Genetic aspects ; Glycine - genetics ; Methodology ; Methods ; mutagenesis ; Mutation ; non-coding RNA ; Observations ; Oryza sativa ; Physiological aspects ; Plant Proteins - genetics ; Plants, Genetically Modified - genetics ; Plasmids ; rice ; RNA sequencing ; RNA, Transfer - genetics ; synthetic genes ; Transfer RNA ; Zea mays ; Zea mays - genetics</subject><ispartof>BMC biotechnology, 2016-08, Vol.16 (1), p.58-58, Article 58</ispartof><rights>COPYRIGHT 2016 BioMed Central Ltd.</rights><rights>Copyright BioMed Central 2016</rights><rights>The Author(s). 2016</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c695t-69017c15c28c5b2cb6f48af6827dddaa915c5d07330c0738fb89b1690317804a3</citedby><cites>FETCH-LOGICAL-c695t-69017c15c28c5b2cb6f48af6827dddaa915c5d07330c0738fb89b1690317804a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4982333/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4982333/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27515683$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Qi, Weiwei</creatorcontrib><creatorcontrib>Zhu, Tong</creatorcontrib><creatorcontrib>Tian, Zhongrui</creatorcontrib><creatorcontrib>Li, Chaobin</creatorcontrib><creatorcontrib>Zhang, Wei</creatorcontrib><creatorcontrib>Song, Rentao</creatorcontrib><title>High-efficiency CRISPR/Cas9 multiplex gene editing using the glycine tRNA-processing system-based strategy in maize</title><title>BMC biotechnology</title><addtitle>BMC Biotechnol</addtitle><description>CRISPR/Cas9 genome editing strategy has been applied to a variety of species and the tRNA-processing system has been used to compact multiple gRNAs into one synthetic gene for manipulating multiple genes in rice.
We optimized and introduced the multiplex gene editing strategy based on the tRNA-processing system into maize. Maize glycine-tRNA was selected to design multiple tRNA-gRNA units for the simultaneous production of numerous gRNAs under the control of one maize U6 promoter. We designed three gRNAs for simplex editing and three multiple tRNA-gRNA units for multiplex editing. The results indicate that this system not only increased the number of targeted sites but also enhanced mutagenesis efficiency in maize. Additionally, we propose an advanced sequence selection of gRNA spacers for relatively more efficient and accurate chromosomal fragment deletion, which is important for complete abolishment of gene function especially long non-coding RNAs (lncRNAs). Our results also indicated that up to four tRNA-gRNA units in one expression cassette design can still work in maize.
The examples reported here demonstrate the utility of the tRNA-processing system-based strategy as an efficient multiplex genome editing tool to enhance maize genetic research and breeding.</description><subject>breeding</subject><subject>Clustered Regularly Interspaced Short Palindromic Repeats</subject><subject>Corn</subject><subject>CRISPR-Associated Proteins</subject><subject>gene editing</subject><subject>Gene Editing - methods</subject><subject>Gene mutations</subject><subject>Genes</subject><subject>Genes, Plant - genetics</subject><subject>Genetic aspects</subject><subject>Glycine - genetics</subject><subject>Methodology</subject><subject>Methods</subject><subject>mutagenesis</subject><subject>Mutation</subject><subject>non-coding RNA</subject><subject>Observations</subject><subject>Oryza sativa</subject><subject>Physiological aspects</subject><subject>Plant Proteins - genetics</subject><subject>Plants, Genetically Modified - genetics</subject><subject>Plasmids</subject><subject>rice</subject><subject>RNA sequencing</subject><subject>RNA, Transfer - genetics</subject><subject>synthetic genes</subject><subject>Transfer RNA</subject><subject>Zea mays</subject><subject>Zea mays - genetics</subject><issn>1472-6750</issn><issn>1472-6750</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>KPI</sourceid><sourceid>BENPR</sourceid><recordid>eNqNks1u1DAUhS1ERduBB2CDIrGhi7S2EzvOBmk0onTUilZTYGs5zk3GVeJMY6dqeHocppQOQqK2_CPf71xd2wehtwQfEyL4iSNU5DzGJIywi-kLdEDSjMY8Y_jlk_0-OnTuBmOSCcxfoX2aMcK4SA6QOzP1OoaqMtqA1WO0WC2vr1YnC-XyqB0abzYN3Ec1WIigNN7YOhrcNPs1RHUzahMifvVlHm_6ToP7FXOj89DGhXJQRs73ykM9RsZGrTI_4DXaq1Tj4M3DOkPfTj99XZzFF5efl4v5Rax5znzM81CvJkxToVlBdcGrVKiKC5qVZalUHkKsxFmSYB1mURUiL0hQJdM1U5XM0Mdt3s1QtFBqsKGSRm5606p-lJ0ycjdizVrW3Z1Mc0GT0Gbow0OCvrsdwHnZGqehaZSFbnCSYpFiLljK_4sSQUiCMQuFztD7v9CbbuhteImJyjDliRB_qFo1II2tulCinpLKecqFIIIxFqjjf1Chl9Aa3VmoTDjfERztCALj4d7XanBOnl8tn80ur1fPZy-_77Jky-q-c66H6vFLCJaTseXW2DIYW07GljRo3j39y0fFbycnPwFWoe9-</recordid><startdate>20160811</startdate><enddate>20160811</enddate><creator>Qi, Weiwei</creator><creator>Zhu, Tong</creator><creator>Tian, Zhongrui</creator><creator>Li, Chaobin</creator><creator>Zhang, Wei</creator><creator>Song, Rentao</creator><general>BioMed Central Ltd</general><general>BioMed Central</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>KPI</scope><scope>3V.</scope><scope>7QO</scope><scope>7TB</scope><scope>7U5</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>L7M</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20160811</creationdate><title>High-efficiency CRISPR/Cas9 multiplex gene editing using the glycine tRNA-processing system-based strategy in maize</title><author>Qi, Weiwei ; Zhu, Tong ; Tian, Zhongrui ; Li, Chaobin ; Zhang, Wei ; Song, Rentao</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c695t-69017c15c28c5b2cb6f48af6827dddaa915c5d07330c0738fb89b1690317804a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>breeding</topic><topic>Clustered Regularly Interspaced Short Palindromic Repeats</topic><topic>Corn</topic><topic>CRISPR-Associated Proteins</topic><topic>gene editing</topic><topic>Gene Editing - methods</topic><topic>Gene mutations</topic><topic>Genes</topic><topic>Genes, Plant - genetics</topic><topic>Genetic aspects</topic><topic>Glycine - genetics</topic><topic>Methodology</topic><topic>Methods</topic><topic>mutagenesis</topic><topic>Mutation</topic><topic>non-coding RNA</topic><topic>Observations</topic><topic>Oryza sativa</topic><topic>Physiological aspects</topic><topic>Plant Proteins - genetics</topic><topic>Plants, Genetically Modified - genetics</topic><topic>Plasmids</topic><topic>rice</topic><topic>RNA sequencing</topic><topic>RNA, Transfer - genetics</topic><topic>synthetic genes</topic><topic>Transfer RNA</topic><topic>Zea mays</topic><topic>Zea mays - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Qi, Weiwei</creatorcontrib><creatorcontrib>Zhu, Tong</creatorcontrib><creatorcontrib>Tian, Zhongrui</creatorcontrib><creatorcontrib>Li, Chaobin</creatorcontrib><creatorcontrib>Zhang, Wei</creatorcontrib><creatorcontrib>Song, Rentao</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>Gale In Context: Global Issues</collection><collection>ProQuest Central (Corporate)</collection><collection>Biotechnology Research Abstracts</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>BMC biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Qi, Weiwei</au><au>Zhu, Tong</au><au>Tian, Zhongrui</au><au>Li, Chaobin</au><au>Zhang, Wei</au><au>Song, Rentao</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>High-efficiency CRISPR/Cas9 multiplex gene editing using the glycine tRNA-processing system-based strategy in maize</atitle><jtitle>BMC biotechnology</jtitle><addtitle>BMC Biotechnol</addtitle><date>2016-08-11</date><risdate>2016</risdate><volume>16</volume><issue>1</issue><spage>58</spage><epage>58</epage><pages>58-58</pages><artnum>58</artnum><issn>1472-6750</issn><eissn>1472-6750</eissn><abstract>CRISPR/Cas9 genome editing strategy has been applied to a variety of species and the tRNA-processing system has been used to compact multiple gRNAs into one synthetic gene for manipulating multiple genes in rice.
We optimized and introduced the multiplex gene editing strategy based on the tRNA-processing system into maize. Maize glycine-tRNA was selected to design multiple tRNA-gRNA units for the simultaneous production of numerous gRNAs under the control of one maize U6 promoter. We designed three gRNAs for simplex editing and three multiple tRNA-gRNA units for multiplex editing. The results indicate that this system not only increased the number of targeted sites but also enhanced mutagenesis efficiency in maize. Additionally, we propose an advanced sequence selection of gRNA spacers for relatively more efficient and accurate chromosomal fragment deletion, which is important for complete abolishment of gene function especially long non-coding RNAs (lncRNAs). Our results also indicated that up to four tRNA-gRNA units in one expression cassette design can still work in maize.
The examples reported here demonstrate the utility of the tRNA-processing system-based strategy as an efficient multiplex genome editing tool to enhance maize genetic research and breeding.</abstract><cop>England</cop><pub>BioMed Central Ltd</pub><pmid>27515683</pmid><doi>10.1186/s12896-016-0289-2</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | breeding Clustered Regularly Interspaced Short Palindromic Repeats Corn CRISPR-Associated Proteins gene editing Gene Editing - methods Gene mutations Genes Genes, Plant - genetics Genetic aspects Glycine - genetics Methodology Methods mutagenesis Mutation non-coding RNA Observations Oryza sativa Physiological aspects Plant Proteins - genetics Plants, Genetically Modified - genetics Plasmids rice RNA sequencing RNA, Transfer - genetics synthetic genes Transfer RNA Zea mays Zea mays - genetics |
| title | High-efficiency CRISPR/Cas9 multiplex gene editing using the glycine tRNA-processing system-based strategy in maize |
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