Rapid CHO Transcriptome Analysis for Adventitious Agent Detection

Performance Period: 6/1/2020 to 11/30/2021

The goal of this project is to apply existing genomic analysis technologies to identify changes in miRNA expression in CHO that could be used to assay infection by adventitious agents (AA). Our initial work using qPCR has identified several miRNA whose expression levels are significantly impacted under conditions of CHO viral infection. Here we will expand the library of miRNA biomarkers for CHO infection using next-generation sequencing (NGS) methods, both at MilliporeSigma and the RPI Genome Core facility. NGS work will include efforts to identify biomarkers that are specific to a particular AA and those that signal a more general infection. Probes will be constructed against the miRNA in the library so they can be quickly and routinely assayed using micelle-tagging electrophoresis (MTE), a rapid, PCR-less method of nucleic-acid quantitation. In final form, we will deliver a technology that can screen for infections continually during the biomanufacturing process, with a rapid readout time and low incidence of false-positive results.