Targeted Knockout of the dhfr, glul, bak1, and bax Genes by the Multiplex Genome Editing in CHO Cells

Орлова, Н. А.; Dayanova, Lutsia K.; Gayamova, E. A.; Sinegubova, Maria; Ковнир, С. В.; Vorobiev, I. I.

doi:10.1134/s1607672922010082

Cited by 6 publications

(1 citation statement)

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“…Orlova et al simultaneously edited four genes (dhfr, glul, BAX, and BAK1) of CHO S cells with the Cas9 enzyme utilizing one optimal gRNA for each gene in an effort to speed up the selection of modified clones. Four gRNAs may be used to alter the CHO cell genome in a multiplex approach, which allows for knocking out of seven out of eight target alleles (Orlova et al, 2022).…”

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%