An LDHa single allele CHO cell mutant exhibits altered metabolic state and enhanced culture performance
Author
dc.contributor.author
Wilkens, Camila A.
Author
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Vishwanathan, Nandita
Author
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Baltes, Nicholas J.
Author
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Lucero, Alicia T.
Author
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Hu, Wei Shou
Author
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Gerdtzen, Ziomara P.
Admission date
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2019-10-15T12:23:57Z
Available date
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2019-10-15T12:23:57Z
Publication date
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2019
Cita de ítem
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Journal of Chemical Technology and Biotechnology, Volumen 94, Issue 5, 2019, Pages 1488-1498
Identifier
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10974660
Identifier
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02682575
Identifier
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10.1002/jctb.5906
Identifier
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https://repositorio.uchile.cl/handle/2250/171657
Abstract
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BACKGROUND: Reducing lactate production in animal cell cultures has been reported to improve cell culture performance and productivity of recombinant protein. A novel genome editing tool, CRISPR/Cas, has been used widely to induce double-strand breaks in the genome and introduce targeted mutations efficiently. RESULTS: In the present work, we used a publicly available human-codon optimized CRISPR/Cas system to introduce mutations in one of the LDHa gene copies to obtain an LDHa single-allele knockout Chinese hamster ovary (CHO) cell clone to analyze its effect over cell metabolism. Fed-batch cultures were conducted in order to evaluate the culture performance of mutant cells. Results show that cell growth was reduced and metabolism was modified by the LDHa single-allele knockout, whereas specific protein rate and volumetric production were greatly enhanced. Additionally, the first in-depth analysis of the metabolic effects of LDHa single allele knock