CHO Cell Engineering at the NICB
The NICB & Pfizer: an SFI funded collaboration
The CHO cell engineering work at the NICB is currently involved with an SFI funded collaboration. As part of the collaboration between the NICB and Pfizer there is a requirement for functional validation of the gene and protein targets emerging from the profiling studies. As a result of the profiling work a number of genes and proteins have been linked to specific favourable cell phenotypes including survival, growth and protein production. To identify if these genes and proteins are in fact effectors of these phenotypes functional validation using siRNA based gene silencing and cDNA overexpression is carried out and the resulting phenotypic changes are assayed.
MicroRNAs are short, non-coding RNA molecules that regulate gene expression at the post-transcriptional level. There is considerable interest in deciphering their role in various biological systems including in CHO cells. Scientists involved in cell engineering at the NICB have performed miRNA expression profiling on temperature-shifted CHO cells and subsequently isolated and cloned the first CHO miRNA. More recent work has illustrated that manipulating the expression of individual miRNAs in these cells may represent a novel genetic engineering approach to improving recombinant protein production by these cells.
Click here to meet our CHO research group
- Guava EasyCyte Plus benchtop flow cytometer
- AB7500, AB7900HT qRT-PCR instruments
- QX100 digital droplet PCR platform
- Kuhner controlled environment shaker-incubators
- Sartorius-Stedim BIOSTAT 2L bioreactors
- Specialist plasmid vector design and construction
- Affymetrix GeneChip microarry suite
- cDNA overexpression
- microRNA overexpression/depletion
- CRISPR-Cas9 gene deletion
- Inducible systems
- Recombinase-mediated Cassette exchange (RMCE)
- Affymetrix Microarray system
- Agilent Bioanalyser
- Guava Easycyte cell analyser
- BD FACS-Aria cell sorter
- Kelly PS, Gallagher C, Clynes M, Barron N. (2015) Conserved microRNA function as a basis for Chinese hamster ovary cell engineering. Biotechnol Lett. 2015 Apr;37(4):787-98. doi: 10.1007/s10529-014-1751-7. Epub 2014 Dec 21.
- Gammell P., Barron N. and Clynes M. (2007) Initial identification of low temperature and culture stage induction of miRNA expression in suspension CHO-K1 cells. J.Biotechnology. 2007, June (130)213-218
- Gammell P. (2007) MicroRNAs: recently discovered key regulators of Proliferation and Apoptosis in Animal cells. Cytotechnology. 2007 Apr;53(1-3):55-63
- Kumar N, Gammell P and Clynes M. (2007) A summary of recent methods employed and the effects of proliferation control in product secreting CHO cell lines Cytotechnology. 2007 Apr;53(1-3):33-46.
- Gammell P , Barron N, Clynes M. A method of producing recombinant biological products Irish patent filed 4 August 2006