Improved protein expression in HEK293 cells by over-expressing miR-22 and knocking-out its target gene, HIPK1

Sarah Inwood; Laura Abaandou; Michael Betenbaugh; Joseph Shiloach

doi:10.1016/j.nbt.2019.08.004

Improved protein expression in HEK293 cells by over-expressing miR-22 and knocking-out its target gene, HIPK1

N Biotechnol. 2020 Jan 25:54:28-33. doi: 10.1016/j.nbt.2019.08.004. Epub 2019 Aug 16.

Authors

Sarah Inwood¹, Laura Abaandou², Michael Betenbaugh³, Joseph Shiloach⁴

Affiliations

¹ Biotechnology Core Laboratory NIDDK, NIH, Bethesda, Maryland, 20892, USA; Department of Chemical and Biomolecular Engineering Johns Hopkins University, Baltimore, Maryland, 21218, USA.
² Biotechnology Core Laboratory NIDDK, NIH, Bethesda, Maryland, 20892, USA.
³ Department of Chemical and Biomolecular Engineering Johns Hopkins University, Baltimore, Maryland, 21218, USA.
⁴ Biotechnology Core Laboratory NIDDK, NIH, Bethesda, Maryland, 20892, USA. Electronic address: Josephs@niddk.nih.gov.

Abstract

Stable cell lines can continuously produce a recombinant protein without the need to repeatedly engineer the genome. In a previous study HIPK1, Homeodomain-interacting Protein Kinase 1, was found to be a target of the microRNA miR-22 that, when repressed, improved expression of both an intracellular and a secreted protein. In this report, HEK293 cells stably over-expressing miR-22 were compared with HEK293 with knockout of HIPK1, executed by CRISPR/Cas9, for their ability to improve recombinant protein expression. In this model case of luciferase, over-expression of miR-22 improved overall activity 2.4-fold while the HIPK1 knockout improved overall activity 4.7-fold.

Keywords: CRISPR/Cas9; HIPK1; Protein expression; Stable; miR-22.

MeSH terms

Gene Expression
Gene Knockout Techniques
HEK293 Cells
Humans
MicroRNAs / genetics*
Protein Biosynthesis / genetics*
Protein Serine-Threonine Kinases / genetics*

Substances

MIRN22 microRNA, human
MicroRNAs
HIPK1 protein, human
Protein Serine-Threonine Kinases