DC Field | Value | Language |
---|---|---|
dc.contributor.author | Lee, namil | ko |
dc.contributor.author | Shin, Jongoh | ko |
dc.contributor.author | Park, Jin-Hyoung | ko |
dc.contributor.author | Lee, Gyun Min | ko |
dc.contributor.author | Suhyung Cho | ko |
dc.contributor.author | Cho, Byung-Kwan | ko |
dc.date.accessioned | 2017-01-23T02:56:36Z | - |
dc.date.available | 2017-01-23T02:56:36Z | - |
dc.date.created | 2017-01-04 | - |
dc.date.created | 2017-01-04 | - |
dc.date.created | 2017-01-04 | - |
dc.date.issued | 2016-11 | - |
dc.identifier.citation | ACS SYNTHETIC BIOLOGY, v.5, no.11, pp.1211 - 1219 | - |
dc.identifier.issn | 2161-5063 | - |
dc.identifier.uri | http://hdl.handle.net/10203/220199 | - |
dc.description.abstract | Chinese hamster ovary (CHO) cells are the preferred host for the production of a wide array of biopharmaceuticals. Thus, efficient and rational CHO cell line engineering methods have been in high demand to improve quality and productivity. Here, we provide a novel genome engineering platform for increasing desirable phenotypes of CHO cells based upon the integrative protocol of high-throughput RNA sequencing and DNA-free RNA guided Cas9 (CRISPR associated protein9) nuclease-based genome editing. For commercial production of therapeutic proteins, CHO cells have been adapted for suspension culture in serum-free media, which is highly beneficial with respect to productivity and economics. To engineer CHO cells for rapid adaptation to a suspension culture, we exploited strand-specific RNA-seq to identify genes differentially expressed according to their adaptation trajectory in serum-free media. More than 180 million sequencing reads were generated and mapped to the currently available 109,152 scaffolds of the CHO-K1 genome. We identified significantly downregulated genes according to the adaptation trajectory and then verified their effects using the genome editing method. Growth-based screening and targeted amplicon sequencing revealed that the functional deletions of Igfbp4 and AqpI gene accelerate suspension adaptation of CHO-K1 cells. The availability of this strand-specific transcriptome sequencing and DNA-free RNA-guided Cas9 nuclease mediated genome editing facilitates the rational design of the CHO cell genome for efficient production of high quality therapeutic proteins. | - |
dc.language | English | - |
dc.publisher | AMER CHEMICAL SOC | - |
dc.subject | HAMSTER OVARY CELLS | - |
dc.subject | SERUM-FREE | - |
dc.subject | RECOMBINANT PROTEINS | - |
dc.subject | GENOME | - |
dc.subject | EXPRESSION | - |
dc.subject | LINE | - |
dc.subject | TRANSCRIPTOME | - |
dc.subject | RIBONUCLEOPROTEINS | - |
dc.subject | INTERFERENCE | - |
dc.subject | INACTIVATION | - |
dc.title | Targeted Gene Deletion Using DNA-Free RNA-Guided Cas9 Nuclease Accelerates Adaptation of CHO Cells to Suspension Culture | - |
dc.type | Article | - |
dc.identifier.wosid | 000388428100004 | - |
dc.identifier.scopusid | 2-s2.0-85017350101 | - |
dc.type.rims | ART | - |
dc.citation.volume | 5 | - |
dc.citation.issue | 11 | - |
dc.citation.beginningpage | 1211 | - |
dc.citation.endingpage | 1219 | - |
dc.citation.publicationname | ACS SYNTHETIC BIOLOGY | - |
dc.identifier.doi | 10.1021/acssynbio.5b00249 | - |
dc.contributor.localauthor | Lee, Gyun Min | - |
dc.contributor.localauthor | Cho, Byung-Kwan | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | Chinese hamster ovary (CHO) cells | - |
dc.subject.keywordAuthor | high-throughput RNA-seq | - |
dc.subject.keywordAuthor | DNA-free CRISPR/Cas9 | - |
dc.subject.keywordAuthor | genome-editing | - |
dc.subject.keywordAuthor | suspension culture | - |
dc.subject.keywordPlus | HAMSTER OVARY CELLS | - |
dc.subject.keywordPlus | SERUM-FREE | - |
dc.subject.keywordPlus | RECOMBINANT PROTEINS | - |
dc.subject.keywordPlus | GENOME | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | LINE | - |
dc.subject.keywordPlus | TRANSCRIPTOME | - |
dc.subject.keywordPlus | RIBONUCLEOPROTEINS | - |
dc.subject.keywordPlus | INTERFERENCE | - |
dc.subject.keywordPlus | INACTIVATION | - |
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