VP of Biomarker and BioAnalytical Sciences Sangamo Therapeutics Richmond
In vivo genome regulation is an emerging therapeutic field that aims to modulate the expression of genes involved in a variety of human diseases. Sangamo has developed a zinc finger-transcription factor (ZF-TF) platform where ZF-TFs can be engineered to specifically bind to any genomic sequence and modulate expression of a target gene via the transcriptional regulatory domain. To support pharmacology and GLP toxicology studies in cynomolgus monkeys, two reverse transcription-quantitative polymerase chain reaction (RT-qPCR) methods were developed and qualified to measure tissue mRNAs of ZF-TF and target gene expression. Absolute copy number of ZF-TF mRNA was quantified using a standard curve prepared from in vitro transcribed mRNA. Change in expression of the target gene was assessed using a relative quantification method. This presentation discusses challenges of method development, generation of reference control material, and establishment of method qualification criteria.
Learning Objectives:
The audience will understand how Sangamo’s ZF-TF technology is used to modulate gene expression.
The audience will understand how to generate reference materials for both absolute and relative mRNA measurement.
The audience will be able to describe the general process for development and qualification of an RT-qPCR assay for absolute and relative measurement of mRNA in support of animal studies.
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