Reporter Assays

reporter gene assays

used to study

1. signaling pathways

2. gene regulation

3. structure of regulatory elements

simplest form reporter assay

consists of

1. regulatory element of interest cloned into a vector

2. with a reporter gene

reporter gene

encodes a protein with an easily measurable activity (bioluminiesce

5’ (upstream) regulatory elements

affect expression of reporter gene

1. potential promoters

2. known promoters

3. partial promoters

4. enhancer elements

reporter assays can also be used

to study elements in 3’ untranslated region

microRNA recognition elements (MREs)

next step

1. transfecting cell

2. when studying upstream elements, mRNA is transcribed from active promoters

3. experimental conditions can modulate activity of promoter

4. weak promotors make less mrna, strong more

5. mrna translated into protein

6. amount of protein is dependent on activity of regulatory element