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PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing

Author

Listed:
  • Abba Malina

    (McGill University)

  • Christopher J. F. Cameron

    (McGill Centre for Bioinformatics and School of Computer Science, McGill University)

  • Francis Robert

    (McGill University)

  • Mathieu Blanchette

    (McGill Centre for Bioinformatics and School of Computer Science, McGill University)

  • Josée Dostie

    (McGill University
    The Rosalind and Morris Goodman Cancer Research Centre, McGill University)

  • Jerry Pelletier

    (McGill University
    The Rosalind and Morris Goodman Cancer Research Centre, McGill University
    McGill University)

Abstract

In CRISPR-Cas9 genome editing, the underlying principles for selecting guide RNA (gRNA) sequences that would ensure for efficient target site modification remain poorly understood. Here we show that target sites harbouring multiple protospacer adjacent motifs (PAMs) are refractory to Cas9-mediated repair in situ. Thus we refine which substrates should be avoided in gRNA design, implicating PAM density as a novel sequence-specific feature that inhibits in vivo Cas9-driven DNA modification.

Suggested Citation

  • Abba Malina & Christopher J. F. Cameron & Francis Robert & Mathieu Blanchette & Josée Dostie & Jerry Pelletier, 2015. "PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing," Nature Communications, Nature, vol. 6(1), pages 1-6, December.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms10124
    DOI: 10.1038/ncomms10124
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