DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, Hyeran | ko |
dc.contributor.author | Kim, Sang-Tae | ko |
dc.contributor.author | Ryu, Jahee | ko |
dc.contributor.author | Choi, Min Kyung | ko |
dc.contributor.author | Kweon, Jiyeon | ko |
dc.contributor.author | Kang, Beum-Chang | ko |
dc.contributor.author | Ahn, Hyo-Min | ko |
dc.contributor.author | Bae, Suji | ko |
dc.contributor.author | Kim, Jungeun | ko |
dc.contributor.author | Kim, Jin-Soo | ko |
dc.contributor.author | Kim, Sang-Gyu | ko |
dc.date.accessioned | 2017-11-08T05:47:13Z | - |
dc.date.available | 2017-11-08T05:47:13Z | - |
dc.date.created | 2017-11-07 | - |
dc.date.created | 2017-11-07 | - |
dc.date.issued | 2016-08 | - |
dc.identifier.citation | JOURNAL OF INTEGRATIVE PLANT BIOLOGY, v.58, no.8, pp.705 - 712 | - |
dc.identifier.issn | 1672-9072 | - |
dc.identifier.uri | http://hdl.handle.net/10203/226940 | - |
dc.description.abstract | CRISPR-Cas9 system is now widely used to edit a target genome in animals and plants. Cas9 protein derived from Streptococcus pyogenes (SpCas9) cleaves double-stranded DNA targeted by a chimeric single-guide RNA (sgRNA). For plant genome editing, Agrobacterium-mediated T-DNA transformation has been broadly used to express Cas9 proteins and sgRNAs under the control of CaMV 35S and U6/U3 promoter, respectively. We here developed a simple and high-throughput binary vector system to clone a 19-20 bp of sgRNA, which binds to the reverse complement of a target locus, in a large T-DNA binary vector containing an SpCas9 expressing cassette. Two-step cloning procedures: (1) annealing two target-specific oligonucleotides with overhangs specific to the AarI restriction enzyme site of the binary vector; and (2) ligating the annealed oligonucleotides into the two AarI sites of the vector, facilitate the high-throughput production of the positive clones. In addition, Cas9-coding sequence and U6/U3 promoter can be easily exchanged via the Gateway(TM) system and unique EcoRI/XhoI sites on the vector, respectively. We examined the mutation ratio and patterns when we transformed these constructs into Arabidopsis thaliana and a wild tobacco, Nicotiana attenuata. Our vector system will be useful to generate targeted large-scale knock-out lines of model as well as non-model plant. | - |
dc.language | English | - |
dc.publisher | WILEY-BLACKWELL | - |
dc.subject | RNA-GUIDED ENDONUCLEASE | - |
dc.subject | AGROBACTERIUM-MEDIATED TRANSFORMATION | - |
dc.subject | TARGETED MUTAGENESIS | - |
dc.subject | HUMAN-CELLS | - |
dc.subject | NUCLEASES | - |
dc.subject | MULTIPLEX | - |
dc.subject | ARABIDOPSIS | - |
dc.subject | SPECIFICITY | - |
dc.subject | EXPRESSION | - |
dc.subject | MODEL | - |
dc.title | A simple, flexible and high-throughput cloning system for plant genome editing via CRISPR-Cas system | - |
dc.type | Article | - |
dc.identifier.wosid | 000382544800003 | - |
dc.identifier.scopusid | 2-s2.0-85027950432 | - |
dc.type.rims | ART | - |
dc.citation.volume | 58 | - |
dc.citation.issue | 8 | - |
dc.citation.beginningpage | 705 | - |
dc.citation.endingpage | 712 | - |
dc.citation.publicationname | JOURNAL OF INTEGRATIVE PLANT BIOLOGY | - |
dc.identifier.doi | 10.1111/jipb.12474 | - |
dc.contributor.localauthor | Kim, Sang-Gyu | - |
dc.contributor.nonIdAuthor | Kim, Hyeran | - |
dc.contributor.nonIdAuthor | Kim, Sang-Tae | - |
dc.contributor.nonIdAuthor | Ryu, Jahee | - |
dc.contributor.nonIdAuthor | Choi, Min Kyung | - |
dc.contributor.nonIdAuthor | Kweon, Jiyeon | - |
dc.contributor.nonIdAuthor | Kang, Beum-Chang | - |
dc.contributor.nonIdAuthor | Ahn, Hyo-Min | - |
dc.contributor.nonIdAuthor | Bae, Suji | - |
dc.contributor.nonIdAuthor | Kim, Jungeun | - |
dc.contributor.nonIdAuthor | Kim, Jin-Soo | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | AarI-mediated sgRNA cloning | - |
dc.subject.keywordAuthor | CRISPR-Cas9 T-DNA binary vector | - |
dc.subject.keywordAuthor | Exchangeable U6 | - |
dc.subject.keywordAuthor | U3 promoter | - |
dc.subject.keywordAuthor | Gateway compatible Cas9 cloning | - |
dc.subject.keywordPlus | RNA-GUIDED ENDONUCLEASE | - |
dc.subject.keywordPlus | AGROBACTERIUM-MEDIATED TRANSFORMATION | - |
dc.subject.keywordPlus | TARGETED MUTAGENESIS | - |
dc.subject.keywordPlus | HUMAN-CELLS | - |
dc.subject.keywordPlus | NUCLEASES | - |
dc.subject.keywordPlus | MULTIPLEX | - |
dc.subject.keywordPlus | ARABIDOPSIS | - |
dc.subject.keywordPlus | SPECIFICITY | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | MODEL | - |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.