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Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9
Published on: April 10, 2018
RAPID-DASH: fast and efficient assembly of guide RNA arrays for multiplexed CRISPR-Cas9 applications
Asfar Lathif Salaudeen1, Nicholas Mateyko1, Carl G de Boer2
1Genome Science and Technology Graduate Program, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada.
Abstract:
Guide RNA (gRNA) arrays can enable targeting multiple genomic loci simultaneously using CRISPR-Cas9. In this study, we present a streamlined and efficient method to rapidly construct gRNA arrays with up to 10 gRNA units in a single day. We demonstrate that gRNA arrays maintain robust functional activity across all positions, and can incorporate libraries of gRNAs, combining scalability and multiplexing. Our approach will streamline combinatorial perturbation research by enabling the economical and rapid construction, testing, and iteration of gRNA arrays. To facilitate the adoption of this approach, we have made a web tool to design oligo sequences necessary to assemble gRNA arrays.
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