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Methods Mol Biol. 2018;1755:107-120. doi: 10.1007/978-1-4939-7724-6_8.

Reporter Gene Assays Using Transfectable Functional Genomics Libraries.

Author information

1
Department of Genomics, The Genomics Institute of the Novartis Research Foundation, San Diego, CA, USA.
2
Functional Genomics Screening Team, The Genomics Institute of the Novartis Research Foundation, San Diego, CA, USA.
3
Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, USA.
4
California NanoSystems Institute, University of California Los Angeles, Los Angeles, CA, USA.
5
Johnsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, USA.
6
Department of Genomics, The Genomics Institute of the Novartis Research Foundation, San Diego, CA, USA. miraglia@gnf.org.
7
Functional Genomics Screening Team, The Genomics Institute of the Novartis Research Foundation, San Diego, CA, USA. miraglia@gnf.org.

Abstract

Transfectable functional genomics libraries are traditionally the workhorses of functional genomics screening using reporter gene assays. These libraries offer insight into fundamental cellular processes governing health and disease and can be utilized in an arrayed fashion which makes them uniquely suited to deconvolute complicated disease phenotypes and dissect biological networks that would otherwise be inaccessible. Here we give an overview of the principles for the generation, screening and data analysis of such arrayed libraries. Specifically we cover the differences between the various transfectable reagents, library selection and handling, and data analysis to offer a comprehensive understanding of these important technologies and how to apply them.

KEYWORDS:

Automation; Data analysis; Functional genomics; HTS; High-throughput screening; Libraries; Transfection; cDNA; miRNA; siRNA

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