Biology Reference
In-Depth Information
CHAPTER 4
RNAi Methods and Screening: RNAi
Based High-Throughput Genetic
Interaction Screening
Patricia G. Cipriani
*
,
y
and Fabio Piano
*
,
y
,
z
*
Center for Genomics and Systems Biology
y
Department of Biology, New York University, New York, NY 10003
z
New York University Abu Dhabi, Abu Dhabi, United Arab Emirates
Abstract
I. Introduction: Large-Scale RNAi Screening in C. elegans
II. Methods
A. Large-Scale RNAi Screening
B. Scoring of Interactions/Phenotypes
C. Quantification of Embryonic Lethality or Survival on Agar Plates
III. Materials
A. Reagents
B. Buffers and Media
C. Supplies and Equipment
Acknowledgments
References
Abstract
Expanding on decades of mutational analyses, numerous genome-scale RNAi
screens have now been performed in C. elegans, leading to estimates that the
majority of genes with essential functions that can be revealed by single-gene
perturbations have already been identified in this organism. To build on this basic
foundation and uncover condition-dependent or combinatorial effects of non-essen-
tial genes will require even higher-scale screening. Here we describe a method for
performing high-throughput RNAi-based screens in C. elegans in liquid in 96-well
plates, and we explain how to systematically test
for enhancement and
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