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. 2021 Jun 21;11:12969. doi: 10.1038/s41598-021-92320-x

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Overview of RMCE and plasmid vectors for the generation of stably transfected CHO cell pools that co-express IgG and human glycosyltransferase genes. (A) Schematic representation of RMCE. (B) Schematic representation of targeting plasmid vectors carrying IgG rituximab LC and HC genes together with one or two human glycosyltransferase genes and DsRed gene. (C) Schematic representation of plasmid vectors expressing B4GalT1 and ST6Gal1 individually or in combination for random integration. RMCE, recombinase-mediated-cassette-exchange; MCL, master cell line; GOI, gene of interest; ChiP, a chimeric promoter consisting of murine cytomegalovirus (CMV) enhancer, human CMV core promoter and human CMV intron A; mCMV, murine CMV enhancer and promoter; FRT and FRT3, wild-type and mutated flippase recognition targets; Flpe, enhanced recombinase flippase; IRES, wild-type encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES); pA, simian virus 40 polyadenylation signal; LC, light chain cDNA; HC, heavy chain cDNA; G.E.1, glycosyltransferase gene 1; G.E.2, glycosyltransferase gene 2; DsRed, cDNA encoding red fluorescent protein; (ATG-)Puro, start-codon ATG-deleted puromycin resistence gene; Bla, blasticidin-S resistance gene; B4GalT1, β-1,4-Galactosyltransferase 1 gene; ST6Gal1, β-Galactoside -α-2,6-sialyltransferase 1 gene.