Hi Pam, In the "loppin" example, if you found that your gene of interest can be expressed in both female and male cell lines, what will you do? Q1: "can be expressed in both" implies the universal Enhancer is present(since the gene is not normally expressed in male). Will you try to get rid of that universal Enhancer by Bashing?
we are not sure that a 'universal enhancer': all we can say is that we have 'caught' an enhancer that functions, and make our gene expressed in both male and female cell lines. We can take two approaches in parallel: on the one hand, as you say, we can start 'bashing', but we will do so by removing adjacent bits of DNA starting from the extremities of our potential regulatory regions. If the reason why we get expression also in the male cells is that we have 'caught' an additional regulatory region, that does not normally act on our gene of interest, this strategy will take care of it, as we will eventually remove it.
At the same time (on separate samples) we can try to extend our potential regulatory region. It is very possible that the reason why we get expression in both cell lines is that we are missing a regulatory region/element that is essential for keeping the gene off in male cells. By extending our potential regulatory region, we should eventually 'catch' it.
Hi Pam,
ReplyDeleteIn the "loppin" example, if you found that your gene of interest can be expressed in both female and male cell lines, what will you do?
Q1: "can be expressed in both" implies the universal Enhancer is present(since the gene is not normally expressed in male). Will you try to get rid of that universal Enhancer by Bashing?
Best,
cindy
Hi Cindy,
ReplyDeletewe are not sure that a 'universal enhancer': all we can say is that we have 'caught' an enhancer that functions, and make our gene expressed in both male and female cell lines. We can take two approaches in parallel: on the one hand, as you say, we can start 'bashing', but we will do so by removing adjacent bits of DNA starting from the extremities of our potential regulatory regions. If the reason why we get expression also in the male cells is that we have 'caught' an additional regulatory region, that does not normally act on our gene of interest, this strategy will take care of it, as we will eventually remove it.
At the same time (on separate samples) we can try to extend our potential regulatory region. It is very possible that the reason why we get expression in both cell lines is that we are missing a regulatory region/element that is essential for keeping the gene off in male cells. By extending our potential regulatory region, we should eventually 'catch' it.
Cheers
Pam
That explains everything. Thanks, Pam. =)
ReplyDeleteCindy