Question

You have just been hired in Dr. Nowotarski's research lab where they are studying the effects an RNA binding protein named TTP has on ornithine decarboxylase mRNA stability. You are given a cell line that is devoid of TTP and want to decipher what happens to the ODC mRNA when you add TTP back into these cells. Furthermore, you know the sequence of mouse TTP and have a pcDNA 3.1 empty vector. How would you go about conducting an experiment in which you add TTP back into these cells?

Answer 1

To add TTP back into the TTP-deficient cells, you can clone the TTP gene into a vector and transfect the cells with the vector containing the TTP gene. Then, analyze the effects of added TTP by examining the stability of the ODC mRNA.

Step-by-step explanation:

To conduct an experiment in which you add TTP back into the TTP-deficient cells, you would need to introduce the TTP gene into the cells using a suitable vector. Since you have the sequence of mouse TTP and an empty pcDNA 3.1 vector, you can clone the TTP gene into the vector using molecular biology techniques.

Once you have the TTP gene cloned into the vector, you would then transfect the TTP-deficient cells with the pcDNA 3.1 vector containing the TTP gene. This can be done using methods such as lipid-mediated transfection or electroporation.

After transfection, you can analyze the effects of adding TTP back into the cells by examining the stability of the ODC mRNA. You can use techniques such as qPCR or RNA stability assays to compare the stability of ODC mRNA in cells with and without TTP.