Enzymatically Generated CRISPR Libraries for Genome Labeling and Screening

Andrew B Lane; Magdalena Strzelecka; Andreas Ettinger; Andrew W Grenfell; Torsten Wittmann; Rebecca Heald

doi:10.1016/j.devcel.2015.06.003

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.

Authors

Andrew B Lane¹, Magdalena Strzelecka², Andreas Ettinger³, Andrew W Grenfell², Torsten Wittmann³, Rebecca Heald⁴

Affiliations

¹ Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA 94720-3200, USA. Electronic address: andylane@berkeley.edu.
² Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA 94720-3200, USA.
³ Department of Cell & Tissue Biology, University of California at San Francisco, San Francisco, CA 94143-0512, USA.
⁴ Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA 94720-3200, USA. Electronic address: bheald@berkeley.edu.

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.

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

Substances

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

Abstract

Publication types

MeSH terms

Substances

Grants and funding