Ac/Ds transposition for CRISPR/dCas9-SID4x epigenome modulation in zebrafish

Author:

Chong-Morrison Vanessa1ORCID,Mayes Sarah1ORCID,Simões Filipa C.12ORCID,Senanayake Upeka1ORCID,Carroll Dervla S.1ORCID,Riley Paul R.2ORCID,Wilson Stephen W.3ORCID,Sauka-Spengler Tatjana14ORCID

Affiliation:

1. University of Oxford, Weatherall Institute of Molecular Medicine 1 , Radcliffe Department of Medicine, Oxford OX3 9DS , UK

2. University of Oxford, Institute of Developmental and Regenerative Medicine 2 , Department of Physiology, Anatomy and Genetics, Oxford OX3 7DQ , UK

3. University College London, Department of Cell & Developmental Biology 3 , London WC1E 6BT , UK

4. Stowers Institute for Medical Research 4 , Kansas City, MO 64110 , USA

Abstract

ABSTRACT Due to its genetic amenability coupled with advances in genome editing, zebrafish is an excellent model to examine the function of (epi)genomic elements. Here, we repurposed the Ac/Ds maize transposition system to efficiently characterise zebrafish cis-regulated elements, also known as enhancers, in F0-microinjected embryos. We further used the system to stably express guide RNAs enabling CRISPR/dCas9-interference (CRISPRi) perturbation of enhancers without disrupting the underlying genetic sequence. In addition, we probed the phenomenon of antisense transcription at two neural crest gene loci. Our study highlights the utility of Ac/Ds transposition as a new tool for transient epigenome modulation in zebrafish.

Funder

Wellcome Trust

RDM Pump Priming

Clarendon Fund Fellowship

BHF Intermediate Basic Science Research Fellowship

BHF Programme Grant and Chair Award

Publisher

The Company of Biologists

Subject

General Agricultural and Biological Sciences,General Biochemistry, Genetics and Molecular Biology

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