Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo

Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo

Summary: Here, we describe a highly efficient, medium-throughput strategy for cloning and in vivo screening of putative enhancers using the chick embryo. By incorporating 48 unique nanotags for use in NanoString nCounter® across three different fluorescent reporters and developing a rapid and effici...

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Bibliographic Details
Main Authors: Ruth M. Williams, Tatjana Sauka-Spengler
Format: Article
Language:English
Published: Elsevier 2021-06-01
Series:STAR Protocols
Subjects:
Model Organisms
Molecular Biology
Gene Expression
CRISPR
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721002148
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spelling doaj-31d78f653f76480b86d84bf0336615642021-06-21T04:25:27ZengElsevierSTAR Protocols2666-16672021-06-0122100507Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryoRuth M. Williams0Tatjana Sauka-Spengler1University of Oxford, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, UK; Corresponding authorUniversity of Oxford, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, UK; Corresponding authorSummary: Here, we describe a highly efficient, medium-throughput strategy for cloning and in vivo screening of putative enhancers using the chick embryo. By incorporating 48 unique nanotags for use in NanoString nCounter® across three different fluorescent reporters and developing a rapid and efficient digestion/ligation type IIs restriction enzyme-based cloning protocol, we develop a multiplexed approach for rapidly identifying enhancer activity.For complete details on the use and execution of this protocol, please see Williams et al. (2019).http://www.sciencedirect.com/science/article/pii/S2666166721002148Model OrganismsMolecular BiologyGene ExpressionCRISPR
collection DOAJ
language English
format Article
sources DOAJ
author Ruth M. Williams
Tatjana Sauka-Spengler
spellingShingle Ruth M. Williams
Tatjana Sauka-Spengler
Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
STAR Protocols
Model Organisms
Molecular Biology
Gene Expression
CRISPR
author_facet Ruth M. Williams
Tatjana Sauka-Spengler
author_sort Ruth M. Williams
title Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_short Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_full Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_fullStr Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_full_unstemmed Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_sort rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
publisher Elsevier
series STAR Protocols
issn 2666-1667
publishDate 2021-06-01
description Summary: Here, we describe a highly efficient, medium-throughput strategy for cloning and in vivo screening of putative enhancers using the chick embryo. By incorporating 48 unique nanotags for use in NanoString nCounter® across three different fluorescent reporters and developing a rapid and efficient digestion/ligation type IIs restriction enzyme-based cloning protocol, we develop a multiplexed approach for rapidly identifying enhancer activity.For complete details on the use and execution of this protocol, please see Williams et al. (2019).
topic Model Organisms
Molecular Biology
Gene Expression
CRISPR
url http://www.sciencedirect.com/science/article/pii/S2666166721002148
work_keys_str_mv AT ruthmwilliams rapidandefficientenhancercloningandinvivoscreeningusingthedevelopingchickembryo
AT tatjanasaukaspengler rapidandefficientenhancercloningandinvivoscreeningusingthedevelopingchickembryo
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