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Methods Mol Biol. 2018;1755:121-133. doi: 10.1007/978-1-4939-7724-6_9.

Reporter Gene Assays Using Viral 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

While transfectable libraries are the workhorse for many screening cores, there is one obvious area where these reagents are not useful-hard to transfect cell lines and primary cells. One solution to this problem is the use of virus to introduce genomic reagents. This strategy is more commonplace now than ever before with libraries covering cDNAs, shDNAs, miRNAs, and guide RNAs readily available. Maintenance and use of these libraries are more challenging than the transient transfection approach due to the viral production step, and the infrastructure necessary to generate them. The following pages will delve into the details for working with arrayed well formats for both lentiviral and retroviral libraries.

KEYWORDS:

CRISPR; Functional genomics; Gain of function; Lentiviral; Library management; Loss of function; Retroviral; cDNA; miRNA; shRNA

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