Timecourse RNA Sequencing of three CHO cell lines: non-producing CHOZN®GS-/-, an IgG1 producer, and an EPO-Fc producer
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ABSTRACT:
PROVIDER: PRJNA899862 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR22246900_1.fastq.gz | Fastqsanger.gz | |||
| SRR22246900_2.fastq.gz | Fastqsanger.gz | |||
| SRR22246901_1.fastq.gz | Fastqsanger.gz | |||
| SRR22246901_2.fastq.gz | Fastqsanger.gz | |||
| SRR22246902_1.fastq.gz | Fastqsanger.gz |
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Project description:We analyzed the transcriptomic response in three distinct CHO cell lines: a non-producing host cell line (CHOZN®GS-/-), an immunoglobulin G1 (IgG1) producer, and an erythropoietin Fc fusion (EPO-Fc) producer. We compared the growth and production characteristics of all three cell lines during fed-batch culture. High throughput RNA sequencing (RNASeq) and quantitative polymerase chain reaction (qPCR) were used to study differential gene expression analysis of the timecourse dataset with the host cell line CHOZN®GS-/- as the reference. The objective of this analysis was to study the common and unique responses of cell lines producing different protein products. This analysis showed enrichment and transient upregulation of mRNAs involved in ER stress, the UPR, and oxidative protein folding and processing in both protein producers. Analysis of temporal differential expression profiles led to the identification of novel engineering targets.
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Project description:CD34+ progenitors were isolated from the bone marrow of three healthy volunteers. CD34+CD71+CD45RA- were FACS sorted to enrich for erythroid progenitors. The cells were cultured for four hours with or without EPO in combination with LY294002, and harvested for RNA extraction, amplification and expression analysis. A compound treatment design type is where the response to administration of a compound or chemical (including biological compounds such as hormones) is assayed. Compound Based Treatment: EPO Keywords: compound_treatment_design
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