Chapter title |
Genome-Wide RNAi Screens for RNA Processing Events in Drosophila melanogaster S2 Cells
|
---|---|
Chapter number | 17 |
Book title |
mRNA Processing
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7204-3_17 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7203-6, 978-1-4939-7204-3
|
Authors |
Natoya Peart, Eric J. Wagner |
Abstract |
Over the past 10 years, the design and application of genome-wide screening (GWS) has improved to the point that it can now be done at level of the individual laboratory. The advantages of GWSs compared to classical genetic screens include: immediate identification of a positive scoring gene, relatively short period of time necessary to conduct the screen (as little as 1 week), cell lines do not present developmental needs for gene expression that an organism normally would, and validation/confirmation of results is straightforward. Here, we describe a general protocol for GWS to be conducted in Drosophila melanogaster S2 cells. We provide specific details on what type of experiments must be done before initiating a screen, the materials that are required to conduct a screen, and make suggestions on methods to carry out secondary screening and counter-screening once the initial GWS is complete. Multiple considerations are also raised that focus on how to anticipate false positives/negatives and how to minimize their occurrence through intelligent design. Finally, we provide specific examples of data that our group has gathered from published genome-wide screens in order to exemplify how "hits" are scored and confirmed. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 4 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 1 | 25% |
Student > Postgraduate | 1 | 25% |
Unknown | 2 | 50% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 50% |
Unknown | 2 | 50% |