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Multi-copy targeted integration for accelerated development of high-producing CHO cells.

Sergeeva D, Lee GM, Nielsen LK, Grav LM
ACS Synth Biol. 2020 Aug 24. doi: 10.1021/acssynbio.0c00322. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
154826pJ204-loxP-EF1a-mCherry-NeoR-lox2272-BGHpA-siteA-HDRLanding pad for Cre/lox-based recombinase-mediated cassette exchange (RMCE) in CHO cells, donor plasmid for CRISPR/Cas9-mediated targeted integration in CHO cells (site A)
154827pJ204-loxP-EF1a-tagBFP-HygR-lox2272-BGHpA-siteT9-HDRLanding pad for Cre/lox-based recombinase-mediated cassette exchange (RMCE) in CHO cells, donor plasmid for CRISPR/Cas9-mediated targeted integration in CHO cells (site T9)
154828pJ204-loxP-100RPU.2-coEPO-lox2272Donor plasmid for Cre/lox-based recombinase-mediated cassette exchange (RMCE), expressing one copy of erythropoietin
154829pJ204-loxP-100RPU.2-coEPO-BGHpA-cHS4-100RPU.2-coEPO-lox2272Donor plasmid for Cre/lox-based recombinase-mediated cassette exchange (RMCE), expressing two copies of erythropoietin
154830pJ607-CMV-GFP-2A-Cas9GFP-2A-Cas9 (CHO codon-optimized)
154831pRSFDuet1-sgRNA(siteA)sgRNA for CRISPR/Cas9-mediated targeted integration into genomic site A in CHO cells
154832pRSFDuet1-sgRNA(siteT9)sgRNA for CRISPR/Cas9-mediated targeted integration into genomic site T9 in CHO cells
154833pJ204-loxP-100RPU.2-ETN-lox2272Donor plasmid for Cre/lox-based recombinase-mediated cassette exchange (RMCE), expressing one copy of etanercept
154834pJ204-loxP-100RPU.2-ETN-BGHpA-cHS4-100RPU.2-ETN-lox2272Donor plasmid for Cre/lox-based recombinase-mediated cassette exchange (RMCE), expressing two copies of etanercept

Antibodies from Article