Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq II
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ABSTRACT:
PROVIDER: PRJNA590485 | ENA |
REPOSITORIES: ENA
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Project description:Small-scale bioreactors used in Chinese hamster ovary (CHO) cell line development allow transcriptomic studies on multiple cell lines. Here we define the CHO cell long non-coding RNA (lncRNA) transcriptome from cells grown in controlled miniature bioreactors under fed-batch conditions using RNA-Seq to identify lncRNAs and how the expression of these changes throughout growth and between IgG producers. We identify lncRNAs associated with productivity and growth characteristics, finding that Adapt15, linked to ER stress, GAS5, linked to mTOR signalling/growth arrest, and PVT1, linked to Myc expression, are differentially regulated during fed-batch culture and whose expression relates to productivity and growth. Changes in (non)-coding RNA expression between the seed train and the equivalent day of fed-batch culture are also reported, showing large differences in gene expression between these, and compared with existing datasets. Collectively, we present a comprehensive lncRNA CHO cell profiling and identify targets for engineering growth and productivity characteristics of CHO cells.
2019-11-20 | GSE140669 | GEO
Project description:Small-scale bioreactors used in Chinese hamster ovary (CHO) cell line development allow transcriptomic studies on multiple cell lines. Here we define the CHO cell long non-coding RNA (lncRNA) transcriptome from cells grown in controlled miniature bioreactors under fed-batch conditions using RNA-Seq to identify lncRNAs and how the expression of these changes throughout growth and between IgG producers. We identify lncRNAs associated with productivity and growth characteristics, finding that Adapt15, linked to ER stress, GAS5, linked to mTOR signalling/growth arrest, and PVT1, linked to Myc expression, are differentially regulated during fed-batch culture and whose expression relates to productivity and growth. Changes in (non)-coding RNA expression between the seed train and the equivalent day of fed-batch culture are also reported, showing large differences in gene expression between these, and compared with existing datasets. Collectively, we present a comprehensive lncRNA CHO cell profiling and identify targets for engineering growth and productivity characteristics of CHO cells.
2019-11-20 | GSE140668 | GEO
Project description:Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq
| PRJNA590479 | ENA
Project description:Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq I
| PRJNA590486 | ENA
Project description:How the long non-coding RNA (lncRNA) genome in recombinant protein producing Chinese hamster ovary (CHO) cell lines relates to phenotype is not well described. We therefore defined the CHO cell lncRNA transcriptome from cells grown in controlled miniature bioreactors under fed-batch conditions using RNA-Seq to identify lncRNAs and how the expression of these changes throughout growth and between IgG producers. We identify lncRNAs including Adapt15, linked to ER stress, GAS5, linked to mTOR signaling/growth arrest, and PVT1, linked to Myc expression, which are differentially regulated during fed-batch culture and whose expression correlates to productivity and growth. Changes in (non)-coding RNA expression between the seed train and the equivalent day of fed-batch culture are also reported and compared with existing datasets. Collectively, we present a comprehensive lncRNA CHO cell profiling and identify targets for engineering growth and productivity characteristics of CHO cells.
| S-EPMC6971398 | biostudies-literature
Project description:This SuperSeries is composed of the SubSeries listed below.
2019-11-20 | GSE140671 | GEO
Project description:Biologics have emerged as leading blockbuster therapeutics, but many advanced recombinant protein moieties remain difficult to produce. Here we identify bottlenecks limiting expression of challenging recombinant human proteins through a systems biology analysis of the transcriptomes of CHO and HEK293. Surprisingly, one third of the challenging human proteins displayed an improved secretion upon host cell swapping from CHO to HEK293. While most components of the secretory machinery showed comparable expression levels in both expression hosts, a key set of genes with extreme expression variation was identified. Among these, ATF4 and SRP9 were validated as general productivity boosters in CHO. Further, we found that more heavily glycosylated products benefit more from the elevated activities of the N- and O-glycosyltransferases found in HEK293 cells. Collectively, our results demonstrate the utilization of HEK293 cells for expression rescue of human proteins and suggest a strategy for identification of secretory pathway components improving yield and quality in HEK293 and CHO.
2022-06-22 | GSE157729 | GEO
Project description:In biopharmaceutical production, Chinese hamster ovary (CHO) cells derived from Cricetulus griseus remain the most commonly used host cell for recombinant protein production, especially antibodies. Over the last decade in-depth multi-omics characterization of these CHO cells provided data for extensive cell line engineering and corresponding increases in productivity. exosomes, extracellular vesicles containing proteins and nucleic acids, are barely researched at all in CHO cells. Exosomes have been proven to be an ubiquitous mediator of intercellular communication and are proposed as new biopharmaceutical format for drug delivery, indicator reflecting host cell condition and anti-apoptotic factor in spent media. Here we sequenced non-coding RNA of Exosomes (EXO) and whole cell lysate (WCL) isolated from CHO-K1 Cell Cultures at different growth phases (logarithmic/exponential phase (log/exp), stationary phase (stat), as well as death phase at 80 % viability (80 % ) and 60 % viability (60 %)) via Lexogen Small RNA-Seq Library Prep Kit for Illumina on the Illumina MiSeq platform in PE mode 2 x 36nt.
2021-01-18 | E-MTAB-9894 | biostudies-arrayexpress
Project description:We present the first comprehensive genome-wide view of CHO translational activity during recombinant expression by the application of ribosome profiling. The distribution of translational power in CHO cells was analyzed in the context of the recombinant mRNAs and the endogenous mRNA pool. In an antibody-producing CHO cell line, the recombinant mRNAs were found to be the most abundant transcripts and also sequestered a substantial amount of translating ribosomes (up to 15%). The recombinant mRNAs were translated as efficiently as the endogenous mRNAs, and changes in translation and transcription of the recombinant mRNAs were directly reflected in changes in specific productivity. Interestingly, improvements in bioprocess quality attributes were achieved by depleting the highly expressed and translated inert NeoR mRNA by siRNA-mediated knock-down. This resulted in improved cellular growth, which was accompanied by an 18% increase in antibody titers. This study is the first to carefully map the translatome of the CHO cell to the nucleotide level and to demonstrate how recombinant expression affects the cell on the translational level.
2016-12-21 | GSE79512 | GEO
Project description:Transcriptomic data from CHO-S cells were gathered from multiple time points during batch culture to provide information about gene presence/absence for generation of a CHO-S cell line specific model using the GIMME algorithm.
2016-11-23 | GSE77800 | GEO