title: CORALINA: a universal method for the generation of gRNA libraries for CRISPR-based screening.
abstract: The 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.
  
  
  
  Here 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.
  
  
  
  The 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.
authors.affiliation: Medical Genomics, UCL Cancer Institute, University College London, 72 Huntley Street, London, WC1E 6BT, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Köferle
authors.affiliation: Biostatistics, Institute of Computational Biology, Helmholtz Zentrum, German Research Center for Environmental Health, Ingolstädter Landstraße 1, 85764, Neuherberg, Germany.
authors.forename: Karolina
authors.initials: K
authors.lastname: Worf
authors.affiliation: 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.
authors.forename: Christopher
authors.initials: C
authors.lastname: Breunig
authors.affiliation: 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.
authors.forename: Valentin
authors.initials: V
authors.lastname: Baumann
authors.affiliation: Bill Lyons Informatics Centre, UCL Cancer Institute, University College London, 72 Huntley Street, London, WC1E 6BT, UK.
authors.forename: Javier
authors.initials: J
authors.lastname: Herrero
authors.affiliation: 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.
authors.forename: Maximilian
authors.initials: M
authors.lastname: Wiesbeck
authors.affiliation: Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, England, UK.
authors.forename: Lukas H
authors.initials: LH
authors.lastname: Hutter
authors.affiliation: 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.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Götz
authors.affiliation: Biostatistics, Institute of Computational Biology, Helmholtz Zentrum, German Research Center for Environmental Health, Ingolstädter Landstraße 1, 85764, Neuherberg, Germany.
authors.forename: Christiane
authors.initials: C
authors.lastname: Fuchs
authors.affiliation: Medical Genomics, UCL Cancer Institute, University College London, 72 Huntley Street, London, WC1E 6BT, UK. s.beck@ucl.ac.uk.
authors.forename: Stephan
authors.initials: S
authors.lastname: Beck
authors.affiliation: Epigenetic Engineering, Institute of Stem Cell Research, Helmholtz Zentrum, German Research Center for Environmental Health, Ingolstädter Landstraße 1, 85764, Neuherberg, Germany. stefan.stricker@helmholtz-muenchen.de.|BioMedizinisches Centrum, Ludwig-Maximilian-Universität, Großhaderner Str. 9, 82152, Planegg-Martinsried, Germany. stefan.stricker@helmholtz-muenchen.de.
authors.forename: Stefan H
authors.initials: SH
authors.lastname: Stricker
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D015723:Gene Library; D005818:Genetic Engineering / Q000379:methods; D023281:Genomics / Q000379:methods; D006801:Humans; D051379:Mice; D000094704:RNA, Guide, CRISPR-Cas Systems; D015203:Reproducibility of Results
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems
keywords: Cas9; Elongated protospacer; Epigenetic engineering; Epigenome editing; Genetic engineering; Genome-wide; gRNA library