| Is
Rev-ErbA? mRNA an Antisense Regulator of TR? 2 mRNA Splicing?
Dorothy
Trawick
Marquette
University
Summer
Mentor: Dr. Stephen Munroe
Antisense
regulation involves base pairing between an antisense RNA molecule and
its complementary target RNA resulting in inhibition of target RNA function.
In recent years, many pairs of antisense RNA in mammals have been discovered
through bioinformatic approaches. However, not much is known about the
regulatory function of these RNA antisense pairs. The focus of my project
is to determine if induced transcription of Rev-Erb? mRNA can inhibit expression
of endogenous TR? 2 mRNA.
The
thyroid hormone receptor gene TR? is alternatively spliced into two different
mRNA molecules: TR? 1 and TR? 2. Rev-Erb? mRNA has a 269 nucleotide overlap
with the TR? 2 mRNA, and it is possible that Rev-Erb? effects transcription
of the TR? 2 mRNA through antisense regulation. In order to induce Rev-Erb?
expression, rat adrenal cells (PC12 cells) were transfected with a chimeric
gene that includes the Rev-Erb? sequence and a GFP marker and is regulated
by a tetracycline and doxycycline responsive promoter. Once stable clones
expressing high levels of Rev-Erb? mRNA have been established, RNA can
be collected and the effects of the induced Rev-Erba mRNA on TR? 2 mRNA
levels will be determined though RNase protection assays. |
