Enzymatically Generated CRISPR Libraries for Genome Labeling and Screening

Dev Cell. 2015 Aug 10;34(3):373-8. doi: 10.1016/j.devcel.2015.06.003. Epub 2015 Jul 23.


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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bacterial Proteins*
  • CRISPR-Associated Protein 9
  • Cloning, Molecular
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Endonucleases*
  • Gene Library*
  • Ovum / cytology
  • RNA / genetics*
  • Xenopus


  • Bacterial Proteins
  • RNA
  • CRISPR-Associated Protein 9
  • Cas9 protein, Francisella novicida
  • Endonucleases