Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 90, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/51719
Keywords
Genetics; Issue 90; gene regulation; transcriptional regulation; sequence-activity mapping; reporter assay; library cloning; transfection; tag sequencing; mammalian cells
Categories
Funding
- National Human Genome Research Institute of the National Institutes of Health [R01HG006785]
Ask authors/readers for more resources
The genetic reporter assay is a well-established and powerful tool for dissecting the relationship between DNA sequences and their gene regulatory activities. The potential throughput of this assay has, however, been limited by the need to individually clone and assay the activity of each sequence on interest using protein fluorescence or enzymatic activity as a proxy for regulatory activity. Advances in high-throughput DNA synthesis and sequencing technologies have recently made it possible to overcome these limitations by multiplexing the construction and interrogation of large libraries of reporter constructs. This protocol describes implementation of a Massively Parallel Reporter Assay (MPRA) that allows direct comparison of hundreds of thousands of putative regulatory sequences in a single cell culture dish.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available