DC Field | Value | Language |
---|---|---|
dc.contributor.author | Lee, Jae Seong | ko |
dc.contributor.author | Grav, Lise Marie | ko |
dc.contributor.author | Pedersen, Lasse Ebdrup | ko |
dc.contributor.author | Lee, Gyun Min | ko |
dc.contributor.author | Kildegaard, Helene Faustrup | ko |
dc.date.accessioned | 2016-12-01T08:01:43Z | - |
dc.date.available | 2016-12-01T08:01:43Z | - |
dc.date.created | 2016-11-28 | - |
dc.date.created | 2016-11-28 | - |
dc.date.issued | 2016-11 | - |
dc.identifier.citation | BIOTECHNOLOGY AND BIOENGINEERING, v.113, no.11, pp.2518 - 2523 | - |
dc.identifier.issn | 0006-3592 | - |
dc.identifier.uri | http://hdl.handle.net/10203/214608 | - |
dc.description.abstract | Targeted gene integration into site-specific loci can be achieved in Chinese hamster ovary (CHO) cells via CRISPR/Cas9 genome editing technology and the homology-directed repair (HDR) pathway. The low efficiency of HDR often requires antibiotic selection, which limits targeted integration of multiple genes at multiple sites. To improve HDR-mediated targeted integration, while avoiding the use of selection markers, chemical treatment for increased HDR, and fluorescent enrichment of genome-edited cells was assessed in CHO cells. Chemical treatment did not improve HDR-mediated targeted integration. In contrast, fluorescent markers in Cas9 and donor constructs enable FACS enrichment, resulting in a threefold increase in the number of cells with HDR-mediated genome editing. Combined with this enrichment method, large transgenes encoding model proteins (including an antibody) were successfully targeted integrated. This approach provides a simple and fast strategy for targeted generation of stable CHO production cell lines in a rational way. (C) 2016 Wiley Periodicals, Inc | - |
dc.language | English | - |
dc.publisher | WILEY-BLACKWELL | - |
dc.subject | DNA-REPAIR PATHWAY | - |
dc.subject | MAMMALIAN-CELLS | - |
dc.subject | CHO-CELLS | - |
dc.subject | RECOMBINATION | - |
dc.subject | EFFICIENCY | - |
dc.subject | RAD51 | - |
dc.title | Accelerated Homology-Directed Targeted Integration of Transgenes in Chinese Hamster Ovary Cells Via CRISPR/Cas9 and Fluorescent Enrichment | - |
dc.type | Article | - |
dc.identifier.wosid | 000386755000023 | - |
dc.identifier.scopusid | 2-s2.0-84988926749 | - |
dc.type.rims | ART | - |
dc.citation.volume | 113 | - |
dc.citation.issue | 11 | - |
dc.citation.beginningpage | 2518 | - |
dc.citation.endingpage | 2523 | - |
dc.citation.publicationname | BIOTECHNOLOGY AND BIOENGINEERING | - |
dc.identifier.doi | 10.1002/bit.26002 | - |
dc.contributor.localauthor | Lee, Gyun Min | - |
dc.contributor.nonIdAuthor | Lee, Jae Seong | - |
dc.contributor.nonIdAuthor | Grav, Lise Marie | - |
dc.contributor.nonIdAuthor | Pedersen, Lasse Ebdrup | - |
dc.contributor.nonIdAuthor | Kildegaard, Helene Faustrup | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | Chinese hamster ovary cells | - |
dc.subject.keywordAuthor | CRISPR/Cas9 | - |
dc.subject.keywordAuthor | fluorescent enrichment | - |
dc.subject.keywordAuthor | homology-directed repair | - |
dc.subject.keywordAuthor | targeted integration | - |
dc.subject.keywordPlus | DNA-REPAIR PATHWAY | - |
dc.subject.keywordPlus | MAMMALIAN-CELLS | - |
dc.subject.keywordPlus | CHO-CELLS | - |
dc.subject.keywordPlus | RECOMBINATION | - |
dc.subject.keywordPlus | EFFICIENCY | - |
dc.subject.keywordPlus | RAD51 | - |
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