<scp>CHO</scp>Cell Engineering for Improved Process Performance and Product Quality

Fischer, Simon; Otte, Kerstin

doi:10.1002/9783527811410.ch9

Cited by 7 publications

(4 citation statements)

References 235 publications

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“…Noncoding RNA‐mediated gene silencing 3. Gene KOs (Fischer et al, 2015; Fischer & Otte, 2019). Each of these approaches could target glycosylation genes (Malphettes et al, 2010; Ngantung et al, 2006; Son et al, 2011), cellular metabolism (Chong et al, 2010; Doolan et al, 2010; Jeong et al, 2001; Kim & Lee, 2007; Tabuchi et al, 2010; Zhou et al, 2011), antiapoptotic/proapoptotic pathways (Jeong et al, 2008; J. S. Lee et al, 2013; Wong et al, 2006), cellular productivity (Chung et al, 2004; Jadhav et al, 2012; Romand et al, 2016), cell cycle checkpoint kinases (K. H. Lee et al, 2013), PTMs (Škulj et al, 2014), and so forth.…”

Section: Crispr‐cas9‐mediated Gene Ko For Cell Line Engineeringmentioning

confidence: 99%

CRISPR‐interceded CHO cell line development approaches

Amiri

Adibzadeh

Ghanbari

et al. 2023

Biotech & Bioengineering

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For industrial production of recombinant protein biopharmaceuticals, Chinese hamster ovary (CHO) cells represent the most widely adopted host cell system, owing to their capacity to produce high‐quality biologics with human‐like posttranslational modifications. As opposed to random integration, targeted genome editing in genomic safe harbor sites has offered CHO cell line engineering a new perspective, ensuring production consistency in long‐term culture and high biotherapeutic expression levels. Corresponding the remarkable advancements in knowledge of CRISPR‐Cas systems, the use of CRISPR‐Cas technology along with the donor design strategies has been pushed into increasing novel scenarios in cell line engineering, allowing scientists to modify mammalian genomes such as CHO cell line quickly, readily, and efficiently. Depending on the strategies and production requirements, the gene of interest can also be incorporated at single or multiple loci. This review will give a gist of all the most fundamental recent advancements in CHO cell line development, such as different cell line engineering approaches along with donor design strategies for targeted integration of the desired construct into genomic hot spots, which could ultimately lead to the fast‐track product development process with consistent, improved product yield and quality.

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Section: Crispr‐cas9‐mediated Gene Ko For Cell Line Engineeringmentioning

confidence: 99%

CRISPR‐interceded CHO cell line development approaches

Amiri

Adibzadeh

Ghanbari

et al. 2023

Biotech & Bioengineering

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“…Especially monoclonal antibodies (mAbs), the largest group of Food and Drug Administration‐approved biopharmaceuticals are most commonly produced in CHO cells (Lagassé et al, 2017; Walsh, 2006). Given the high value of these biopharmaceuticals, considerable attention is being paid to methods for accelerating cell line engineering and improving product yields and quality (Fischer & Otte, 2019; Lee et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

“…Similarly, the convenience and simplicity of gene editing have led to the development of multiplex knockouts of disadvantageous genes (Kol et al, 2020). In this way, the CRISPR tool has been used to improve various parameters of CHO cell bioprocessing, such as viability, protein folding, and secretion (Fischer & Otte, 2019; Hansen et al, 2017; Schweickert & Cheng, 2020; Shin et al, 2021). It has also facilitated the control and manipulation of protein glycosylation as well as the elimination of interfering host cell proteins (HCPs) (Amann et al, 2019; Kol et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Expanding the CRISPR toolbox for Chinese hamster ovary cells with comprehensive tools for Mad7 genome editing

Rojek

Basavaraju

Nallapareddy

et al. 2023

Biotech & Bioengineering

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The production of high-value biopharmaceuticals is dominated by mammalian production cells, particularly Chinese hamster ovary (CHO) cells, which have been widely used and preferred in manufacturing processes. The discovery of CRISPR-Cas9 significantly accelerated cell line engineering advances, allowing for production yield and quality improvements. Since then, several other CRISPR systems have become appealing genome editing tools, such as the Cas12a nucleases, which provide broad editing capabilities while utilizing short guide RNAs (gRNAs) that reduce the complexity of the editing systems. One of these is the Mad7 nuclease, which has been shown to efficiently convey targeted gene disruption and insertions in several different organisms. In this study, we demonstrate that Mad7 can generate indels for gene knockout of host cell proteins in CHO cells. We found that the efficiency of Mad7 depends on the addition of protein nuclear localization signals and the gRNAs employed for genome targeting. Moreover, we provide computational tools to design Mad7 gRNAs against any genome of choice and for automated indel detection analysis from next-generation sequencing data. In summary, this paper establishes the application of Mad7 in CHO cells, thereby improving the CRISPR toolbox versatility for research and cell line engineering.

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“…[3] Over the past decade, CHO cell productivity has increased from less than one milligram per liter to grams per liter [4] through process engineering, [5,6] media supplementation, [7,8] and cell line engineering. [9,10] Still, novel targets to engineer CHO cells are desired to increase yield per batch and to provide robust product quality.…”

Section: Introductionmentioning

confidence: 99%

Rosmarinic acid enhances CHO cell productivity and proliferation through activation of the unfolded protein response and the mTOR pathway

Mao,

Schneider,

Robinson

2023

Biotechnology Journal

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Rosmarinic acid (RA) has gained attraction in bioprocessing as a media supplement to improve cellular proliferation and protein production. Here, we observe up to a two‐fold increase in antibody production with RA‐supplementation, and a concentration‐dependent effect of RA on cell proliferation for fed‐batch CHO cell cultures. Contrary to previously reported antioxidant activity, RA increased the reactive oxygen species (ROS) levels, stimulated endoplasmic reticulum (ER) stress, activated the unfolded protein response (UPR), and elicited DNA damage. Despite such stressful events, RA appeared to maintained cell health via mTOR pathway activation; both mTORC1 and mTORC2 were stimulated in RA‐supplemented cultures. By reversing such mTOR pathway activity through either chemical inhibitor addition or siRNA knockdown of genes regulating the mTORC1 and mTORC2 complexes, antibody production, UPR signaling, and stress‐induced DNA damage were reduced. Further, the proliferative effect of RA appeared to be regulated selectively by mTORC2 activation and have reproduced this observation by using the mTORC2 stimulator SC‐79. Analogously, knockdown of mTORC2 strongly reduced XBP1 splicing, which would be expected to reduce antibody folding and secretion, sugging that reduced mTORC2 would correlate with reduced antibody levels. The crosstalk between mTOR activation and UPR upregulation may thus be related directly to the enhanced productivity. Our results show the importance of the mTOR and UPR pathways in increasing antibody productivity, and suggest that RA supplementation may obviate the need for labor‐intensive genetic engineering by directly activating pathways favorable to cell culture performance.This article is protected by copyright. All rights reserved

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CHOCell Engineering for Improved Process Performance and Product Quality

Cited by 7 publications

References 235 publications

CRISPR‐interceded CHO cell line development approaches

CRISPR‐interceded CHO cell line development approaches

Expanding the CRISPR toolbox for Chinese hamster ovary cells with comprehensive tools for Mad7 genome editing

Rosmarinic acid enhances CHO cell productivity and proliferation through activation of the unfolded protein response and the mTOR pathway

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