A comprehensive evaluation of stable expression “hot spot” in the ScltI gene of Chinese hamster ovary cells

Ding, Xuefeng; Chen, Yun; Yang, Zhaoqi; Wu, Hao; Cai, Yanfei; Wang, Enming; Jin, Jian; Li, Huazhong

doi:10.1007/s00253-023-12383-w

Cited by 1 publication

(2 citation statements)

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“…This provides robust evidence for the NC_048595.1 locus' status as a transcriptional hotspot in the CHO cell lines. In a comparable study conducted by Ding et al (2023), it was demonstrated that the exogenous proteins EGFP and HSA were effectively incorporated near the base 1969647 on chromosome NW_003613638.1 of CHO-K1 cells. The resultant monoclonal cell lines, established through 60 consecutive passages, consistently manifested the expression of all associated exogenous proteins, with no notable variations in expression levels detected [23].…”

Section: Discussionmentioning

confidence: 91%

“…In a comparable study conducted by Ding et al (2023), it was demonstrated that the exogenous proteins EGFP and HSA were effectively incorporated near the base 1969647 on chromosome NW_003613638.1 of CHO-K1 cells. The resultant monoclonal cell lines, established through 60 consecutive passages, consistently manifested the expression of all associated exogenous proteins, with no notable variations in expression levels detected [23]. Zhao M et al (2018) employed a similar site-specific integration approach, exploring three sites C12orf35, HPRT, and GRIK1 with potential high transcriptional activities in CHO cells.…”

Section: Discussionmentioning

confidence: 91%

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CRISPR-Cas9 Mediated Stable Expression of Exogenous Proteins in the CHO Cell Line through Site-Specific Integration

Huang,

Habib,

Zhao

et al. 2023

IJMS

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Chinese hamster ovary (CHO) cells are a popular choice in biopharmaceuticals because of their beneficial traits, including high-density suspension culture, safety, and exogenously produced proteins that closely resemble natural proteins. Nevertheless, a decline in the expression of exogenous proteins is noted as culture time progresses. This is a consequence of foreign gene recombination into chromosomes by random integration. The current investigation employs CRISPR-Cas9 technology to integrate foreign genes into a particular chromosomal location for sustained expression. Results demonstrate the successful integration of enhanced green fluorescent protein (EGFP) and human serum albumin (HSA) near base 434814407 on chromosome NC_048595.1 of CHO-K1 cells. Over 60 successive passages, monoclonal cell lines were produced that consistently expressed all relevant external proteins without discernible variation in expression levels. In conclusion, the CHO-K1 cell locus, NC_048595.1, proves an advantageous locus for stable exogenous protein expression. This study provides a viable approach to establishing a CHO cell line capable of enduring reliable exogenous protein expression.

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Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 91%