Safe-Harboring based novel genetic toolkit for Nannochloropsis salina CCMP1776: Efficient overexpression of transgene via CRISPR/Cas9-Mediated Knock-in at the transcriptional hotspot

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dc.contributor.authorRyu, Ae Jinko
dc.contributor.authorJeong, Byeong-ryoolko
dc.contributor.authorKang, Nam Kyuko
dc.contributor.authorJeon, Seungjibko
dc.contributor.authorSohn, Min Giko
dc.contributor.authorYun, Hyo Jinko
dc.contributor.authorLim, Jong Minko
dc.contributor.authorJeong, Seok Wonko
dc.contributor.authorPark, Youn-Ilko
dc.contributor.authorJeong, Won Joongko
dc.contributor.authorPark, Sunghoonko
dc.contributor.authorChang, Yong Keunko
dc.contributor.authorJeong, Ki Junko
dc.date.accessioned2021-09-28T01:10:26Z-
dc.date.available2021-09-28T01:10:26Z-
dc.date.created2021-09-28-
dc.date.created2021-09-28-
dc.date.issued2021-11-
dc.identifier.citationBIORESOURCE TECHNOLOGY, v.340-
dc.identifier.issn0960-8524-
dc.identifier.urihttp://hdl.handle.net/10203/287891-
dc.description.abstractTransgene expression in microalgae can be hampered by transgene silencing and unstable expression due to position effects. To overcome this, "safe harboring" transgene expression system was established for Nanno-chloropsis. Initially, transformants were obtained expressing a sfGFP reporter, followed by screening for high expression of sfGFP with fluorescence-activated cell sorter (FACS). 'T1' transcriptional hotspot was identified from a mutant showing best expression of sfGFP, but did not affect growth or lipid contents. By using a Cas9 editor strain, FAD12 gene, encoding Delta 12-fatty acid desaturase (FAD12), was successfully knocked-in at the T1 locus, resulting in significantly higher expression of FAD12 than those of random integration. Importantly, the "safe harbored" FAD12 transformants showed four-fold higher production of linoleic acid (LA), the product of FAD12, leading to 1.5-fold increase in eicosapentaenoic acid (EPA). This safe harboring principle provide excellent proof of the concept for successful genetic/metabolic engineering of microalgae and other organisms.-
dc.languageEnglish-
dc.publisherELSEVIER SCI LTD-
dc.titleSafe-Harboring based novel genetic toolkit for Nannochloropsis salina CCMP1776: Efficient overexpression of transgene via CRISPR/Cas9-Mediated Knock-in at the transcriptional hotspot-
dc.typeArticle-
dc.identifier.wosid000696313700004-
dc.identifier.scopusid2-s2.0-85112485517-
dc.type.rimsART-
dc.citation.volume340-
dc.citation.publicationnameBIORESOURCE TECHNOLOGY-
dc.identifier.doi10.1016/j.biortech.2021.125676-
dc.contributor.localauthorJeong, Byeong-ryool-
dc.contributor.localauthorChang, Yong Keun-
dc.contributor.localauthorJeong, Ki Jun-
dc.contributor.nonIdAuthorSohn, Min Gi-
dc.contributor.nonIdAuthorYun, Hyo Jin-
dc.contributor.nonIdAuthorLim, Jong Min-
dc.contributor.nonIdAuthorJeong, Seok Won-
dc.contributor.nonIdAuthorPark, Youn-Il-
dc.contributor.nonIdAuthorJeong, Won Joong-
dc.contributor.nonIdAuthorPark, Sunghoon-
dc.description.isOpenAccessN-
dc.type.journalArticleArticle-
dc.subject.keywordAuthorCRISPR/Cas9-
dc.subject.keywordAuthorSafe harboring-
dc.subject.keywordAuthorNannochloropsis salina-
dc.subject.keywordAuthorFatty acid desaturase-
dc.subject.keywordAuthorKnock-in via NHEJ-
dc.subject.keywordPlusCHLAMYDOMONAS-REINHARDTII-
dc.subject.keywordPlusEXPRESSION-
dc.subject.keywordPlusBIOSYNTHESIS-
dc.subject.keywordPlusMICROALGAE-
dc.subject.keywordPlusACYLTRANSFERASES-
dc.subject.keywordPlusSTRAINS-
dc.subject.keywordPlusPROTEIN-
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CBE-Journal Papers(저널논문)
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