Question

1. The gene RGB encodes numerous proteins due to extensive alternative splicing of its 18 exons. One of these proteins, RGB9, plays a crucial role in prenatal development but after development is no longer expressed. Postnatal expression of RGB9 leads to a condition called Button Syndrome, which negatively impacts systemic development. Patients with this disease exhibit neurocognitive impairments and skeletal deformities, among other symptoms.
   1. You decide to create transgenic mice that express a interfering RNA against RGB9. What type of transgenic system do you use and why? There are multiple possible answers.
   2. Draw the gene cassette(s) that will be incorporated into the transgenic mice you described in part a.
   3. You follow the protocol for creating the transgenic mice, but are unsure if the mice have successfully integrated your genetic constructs. What is one way you could check to see if your mice were generated correctly?

Answer 1

a) Transgenic Knockout mice can be used to study the implications of knocking out a specific gene. Also, a transgenic RNAi mice can be used. RNA interference can be used to perform loss-of-function genetic screens.  This can be done either by generating mouse ES cell clones that show RNAi effect and then to use the ES cell clones to produce mice. Another way is by injecting one cell mouse embryos with lentivirus carrying a shRNA- expressing cassette.