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Directed Evolution of Subcloning for CHO

ABSTRACT: Directed Evolution Approach to enhance Efficiency and Speed of Outgrowth during Single Cell Subcloning of Chinese Hamster Ovary Cells

PROVIDER: PRJEB37009 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR3972090.fastq.gz	Fastqsanger.gz
	ERR3972091.fastq.gz	Fastqsanger.gz
	ERR3972092.fastq.gz	Fastqsanger.gz
	ERR3972093.fastq.gz	Fastqsanger.gz
	ERR3972094.fastq.gz	Fastqsanger.gz

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Comparison of transcriptional profile of CHO cells cultured in different media.

Project description:Two clones of Chinese hamster ovary (CHO) cells were cultured in 2 different chemical difined custom media. Transcriptional profile were compeared between clones and media.

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Project description:The effects of changes in bioprocessing parameters (temp, pH, DO) on anti-IL8 IgG1 producing Chinese hamster ovary (CHO) cells were studied on the level of host cell proteome expression using TMT-SPS-MS3.

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A Chinese hamster transcription start site atlas that enables targeted editing of CHO cells

Project description:Gene expression is a key determinant of phenotypes that made Chinese Hamster Ovary (CHO) cells, with their human-like glycosylation profile and high protein titers, one of the most widely used cells for the production of therapeutic proteins and biopharmaceuticals. Engineering CHO gene expression thus holds a key to improve drug quality and cost effective production. However, the success of engineering gene expression or ectopic activation of silent genes to optimize desired pathways requires accurate annotation of the underlying regulatory elements and the transcription start site (TSS). Unfortunately, to date, most TSSs of CHO-expressed genes and the ~50% of hamster genes that are silent in CHO were computationally predicted and are frequently inaccurate. To oust this hurdle, we report revised TSSs annotations for 15,308 Chinese Hamster genes and 4,145 non-coding RNAs based on experimental data from CHO K1 cells and 10 hamster tissues. In the example of the glycosyltransferase gene Mgat3, we further demonstrate how accurate annotations readily facilitate activating silent genes by CRISPRa. Together, we envision that our annotation and data from the Chinese Hamster will provide a rich resource for the CHO community, improve genome engineering efforts and additionally aid comparative and evolutionary studies.

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Cricetulus griseus strain:CHO cell lines

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Project description:Directed biomolecular evolution is widely used to tailor and enhance proteins but has hitherto not been applied in the reprogramming of mammalian cells. Here we describe a novel method to identify artificially enhanced and evolved reprogramming factors by pooled screens with randomised protein libraries, cell selection based on phenotypic readouts and genotyping by amplicon sequencing. We benchmark this approach by identifying artificially enhanced Sox2 and Sox17 factors in pluripotency reprogramming.

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Mechanistic insights into the biological activity of S-Sulfocysteine in CHO cells using a multi-omics approach

Project description:S-Sulfocysteine (SSC), a bioavailable L-cysteine derivative (Cys), is known to be taken up and metabolized in Chinese hamster ovary (CHO) cells used to produce novel therapeutic biological entities. To gain a deeper mechanistic insight into the SSC biological activity and metabolization, a comprehensive multi-omics study was performed on industrially relevant CHO-K1 GS cells throughout a fed-batch process, including metabolomic and proteomic profiling combined with multivariate data and pathway analyses. Multi-layered data and enzymatical assays revealed an intracellular SSC/glutathione mixed disulfide formation and glutaredoxin-mediated reduction, releasing Cys and sulfur species. Increased Cys availability was directed towards glutathione and taurine synthesis, while other Cys catabolic pathways were likewise affected, indicating the cells strive to maintain Cys homeostasis and cellular functions.

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2022-05-20 | PXD027443 | Pride

Cricetulus griseus strain:CHO K1 cell line

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