title: Enzymatically Generated CRISPR Libraries for Genome Labeling and Screening.
abstract: CRISPR-based technologies have emerged as powerful tools to alter genomes and mark chromosomal loci, but an inexpensive method for generating large numbers of RNA guides for whole genome screening and labeling is lacking. Using a method that permits library construction from any source of DNA, we generated guide libraries that label repetitive loci or a single chromosomal locus in Xenopus egg extracts and show that a complex library can target the E. coli genome at high frequency.
authors.affiliation: Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA 94720-3200, USA. Electronic address: andylane@berkeley.edu.
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Lane
authors.affiliation: Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA 94720-3200, USA.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Strzelecka
authors.affiliation: Department of Cell & Tissue Biology, University of California at San Francisco, San Francisco, CA 94143-0512, USA.
authors.forename: Andreas
authors.initials: A
authors.lastname: Ettinger
authors.affiliation: Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA 94720-3200, USA.
authors.forename: Andrew W
authors.initials: AW
authors.lastname: Grenfell
authors.affiliation: Department of Cell & Tissue Biology, University of California at San Francisco, San Francisco, CA 94143-0512, USA.
authors.forename: Torsten
authors.initials: T
authors.lastname: Wittmann
authors.affiliation: Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA 94720-3200, USA. Electronic address: bheald@berkeley.edu.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Heald
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D000076987:CRISPR-Associated Protein 9; D003001:Cloning, Molecular; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004720:Endonucleases; D015723:Gene Library; D010063:Ovum / Q000166:cytology; D012313:RNA / Q000235:genetics*; D014981:Xenopus
chemical_list: D001426:Bacterial Proteins; D012313:RNA; D000076987:CRISPR-Associated Protein 9; C000600049:Cas9 protein, Francisella novicida; D004720:Endonucleases