High-throughput functional analysis of regulatory variants using a massively parallel reporter assay

Summary: Association studies describe genetic associations between noncoding variants and disease susceptibility; however, they do not provide functional insight into the underlying molecular mechanisms of these variants. We present a protocol to assay the regulatory potential of thousands of noncod...

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Bibliographic Details
Main Authors: Kate Delfosse, Chiara Gerhardinger, John L. Rinn, Philipp G. Maass
Format: Article
Language:English
Published: Elsevier 2023-12-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166723006986
Description
Summary:Summary: Association studies describe genetic associations between noncoding variants and disease susceptibility; however, they do not provide functional insight into the underlying molecular mechanisms of these variants. We present a protocol to assay the regulatory potential of thousands of noncoding variants using massively parallel reporter assays. We describe steps for oligo design, generating a plasmid pool, and extracting tag-seq libraries from cells to quantify the tested sequences.For complete details on the use and execution of this protocol, please refer to Oliveros and Delfosse et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667