CORALINA: a universal method for the generation of gRNA libraries for CRISPR-based screening

BMC Genomics

Methodology Article

Javier Herrero¹  Christiane Fuchs²  Karolina Worf²  Lukas H. Hutter³  Christopher Breunig⁴  Valentin Baumann⁴  Maximilian Wiesbeck⁴  Stefan H. Stricker⁴  Stephan Beck⁵  Anna Köferle⁵  Magdalena Götz⁶ 

[1] Bill Lyons Informatics Centre, UCL Cancer Institute, University College London, 72 Huntley Street, WC1E 6BT, London, UK;Biostatistics, Institute of Computational Biology, Helmholtz Zentrum, German Research Center for Environmental Health, Ingolstädter Landstraße 1, 85764, Neuherberg, Germany;Department of Biochemistry, University of Oxford, OX1 3QU, Oxford, England, UK;Epigenetic Engineering, Institute of Stem Cell Research, Helmholtz Zentrum, German Research Center for Environmental Health, Ingolstädter Landstraße 1, 85764, Neuherberg, Germany;BioMedizinisches Centrum, Ludwig-Maximilian-Universität, Großhaderner Str. 9, 82152, Planegg-Martinsried, Germany;Medical Genomics, UCL Cancer Institute, University College London, 72 Huntley Street, WC1E 6BT, London, UK;Neural Stem Cells, Institute of Stem Cell Research, Helmholtz Zentrum, German Research Center for Environmental Health, Ingolstädter Landstraße 1, 85764, Neuherberg, Germany;BioMedizinisches Centrum, Ludwig-Maximilian-Universität, Großhaderner Str. 9, 82152, Planegg-Martinsried, Germany;

关键词: gRNA library;    Genome-wide;    Cas9;    Genetic engineering;    Epigenetic engineering;    Elongated protospacer;    Epigenome editing;   

DOI  :  10.1186/s12864-016-3268-z

 received in 2016-04-08, accepted in 2016-11-05,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundThe bacterial CRISPR system is fast becoming the most popular genetic and epigenetic engineering tool due to its universal applicability and adaptability. The desire to deploy CRISPR-based methods in a large variety of species and contexts has created an urgent need for the development of easy, time- and cost-effective methods enabling large-scale screening approaches.ResultsHere we describe CORALINA (comprehensive gRNA library generation through controlled nuclease activity), a method for the generation of comprehensive gRNA libraries for CRISPR-based screens. CORALINA gRNA libraries can be derived from any source of DNA without the need of complex oligonucleotide synthesis. We show the utility of CORALINA for human and mouse genomic DNA, its reproducibility in covering the most relevant genomic features including regulatory, coding and non-coding sequences and confirm the functionality of CORALINA generated gRNAs.ConclusionsThe simplicity and cost-effectiveness make CORALINA suitable for any experimental system. The unprecedented sequence complexities obtainable with CORALINA libraries are a necessary pre-requisite for less biased large scale genomic and epigenomic screens.

【 授权许可】

CC BY   
© The Author(s). 2016