In Vivo RNAi Agent Services

The cellular RNA interference (RNAi) pathway can be used to effectively regulate gene expression. RNAi has found utility in the drug identification and development pipeline. BIOO Scientific can perform services using a number of different RNAi technologies including synthetic siRNA and vector based approaches to help you facilitate the progress of drug development.

RNAi has catapulted itself into use for therapeutic and animal applications to study gene function on the whole animal system. One of the biggest reasons for moving into animals is current cell culture methods have limitations and are not representative of the true function of a particular gene or process. Tissue culture was first devised at the beginning of the last century as a method for studying the behavior of animal cells free of systemic variations that might arise in the animal both under homeostasis and under stress of an experiment. Although tissue culture has enormous utility, there are many aspects of the characterization done in cultured cells that are not occurring when in the whole organism. For example, cells proliferate in vitro which would not normally in vivo , cell-cell and cell-matrix interactions are reduced because purified cell lines lack heterogeneity and three dimensional architecture found in vivo and the hormonal and nutritional environment is altered. This creates the environment that favors the spreading, migration and proliferation of unspecialized cells rather than the expression of differentiated fractions and increased cellular stress.

One such example of cells with difficulty in adapting to culture are those cells derived from the bronchial respiratory epithelium (Ramirez, Sheridan et al. 2004). Bronchial respiratory epithelium will induce the cyclin dependent kinase inhibitor, p16INK4a and induce growth arrest soon after being placed into a culture dish. Standard cell cultures conditions (20% O2) are harsh. Cells in the body are exposed to oxygen tensions between 1-5% which is far below atmospheric oxygen levels (von Zglinicki 2002). It has been found that this p16INK4a induced growth arrest can be overcome by expressing cdk4, the target of p16. Even with bypassing growth arrest using cdk4 expression, bronchial respiratory epithelium cells will eventually undergo cellular growth arrest due to telomere shortening. The process of telomere-dependant growth arrest, as explained in more detail below, can be overcome by next expressing the catalytic protein component of telomerase (TERT) or through introduction of viral oncogenes (Hahn, Counter et al. 1999). Unfortunately, the example provided by the bronchial epithelium cell experiments discussed above is the norm rather than the exception. Of the over 250 cell types and 10 trillion cells in the human body many, if not most, cannot be grown efficiently in typical culture conditions and those that can, adapt different gene expression patterns than would be typically found in the body.

BIOO offers custom RNAi services in the area of the whole organism. These experiments are typically not easy for most researchers to do as the techniques are very specialized and time consuming to get initiated. We offer a cost effective way of testing specific siRNA molecules in the model system most appropriate for your goals. This will enable researchers to take siRNAs against clinically important targets directly into an animal environment for further validation in an efficient fashion with minimal down time.

In vivo RNAi service workflow:

The complete and integrated RNAi workflow includes the following services:

For additional information on these services, please contact us at RNAi@biooscientific.com or call (512) 707-0921.