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Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq II

ABSTRACT: Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq II

PROVIDER: PRJNA590485 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10493846.fastq.gz	Fastqsanger.gz
	SRR10493846_1.fastq.gz	Fastqsanger.gz
	SRR10493846_2.fastq.gz	Fastqsanger.gz
	SRR10493847.fastq.gz	Fastqsanger.gz
	SRR10493847_1.fastq.gz	Fastqsanger.gz

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Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq II

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.

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Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq I

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Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq

Project description:Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq

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Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq II

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Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq I

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Defining lncRNAs Correlated with CHO Cell Growth and IgG Productivity by RNA-Seq.

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.

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Defining lncRNAs Associated with CHO Cell Growth and IgG Productivity by RNA-Seq

Project description:This SuperSeries is composed of the SubSeries listed below.

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Secretory pathway differences between HEK293 and CHO enable rescue of difficult to express human proteins

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.

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