Modular one-pot assembly of CRISPR arrays enables library generation and reveals factors influencing crRNA biogenesis

Chunyu Liao; Fani Ttofali; Rebecca A. Slotkowski; Steven R. Denny; Taylor D. Cecil; Ryan T. Leenay; Albert J. Keung; Chase L. Beisel

doi:10.1038/s41467-019-10747-3

Nature Communications (Jul 2019)

Modular one-pot assembly of CRISPR arrays enables library generation and reveals factors influencing crRNA biogenesis

Chunyu Liao,
Fani Ttofali,
Rebecca A. Slotkowski,
Steven R. Denny,
Taylor D. Cecil,
Ryan T. Leenay,
Albert J. Keung,
Chase L. Beisel

Affiliations

Chunyu Liao: Department of Chemical and Biomolecular Engineering, North Carolina State University
Fani Ttofali: Department of Chemical and Biomolecular Engineering, North Carolina State University
Rebecca A. Slotkowski: Department of Chemical and Biomolecular Engineering, North Carolina State University
Steven R. Denny: Department of Chemical and Biomolecular Engineering, North Carolina State University
Taylor D. Cecil: Department of Chemical and Biomolecular Engineering, North Carolina State University
Ryan T. Leenay: Department of Chemical and Biomolecular Engineering, North Carolina State University
Albert J. Keung: Department of Chemical and Biomolecular Engineering, North Carolina State University
Chase L. Beisel: Department of Chemical and Biomolecular Engineering, North Carolina State University

DOI: https://doi.org/10.1038/s41467-019-10747-3
Journal volume & issue: Vol. 10, no. 1
pp. 1 – 14

Abstract

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CRISPR array generation is difficult due to reoccurring repeat sequences. Here the authors present CRATES—a modular, one-pot assembly method—and demonstrate the creation of arrays for Cas9, Cas12a and Cas13a for cell-free, bacterial, yeast and mammalian systems.

Published in Nature Communications

ISSN: 2041-1723 (Online)
Publisher: Nature Portfolio
Country of publisher: United Kingdom
LCC subjects: Science
Website: https://www.nature.com/ncomms/

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