Publication

Multiplexed genome engineering by Cas12a and CRISPR arrays encoded on single transcripts

Campa, C. C., Weisbach, N. R., Santinha, A. J., Incarnato, D. & Platt, R. J., Sep-2019, In : Nature Methods. 16, 9, p. 887-893 7 p.

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  • Multiplexed genome engineering by Cas12a and CRISPR arrays encoded

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DOI

  • Carlo C. Campa
  • Niels R. Weisbach
  • António J. Santinha
  • Danny Incarnato
  • Randall J. Platt

The ability to modify multiple genetic elements simultaneously would help to elucidate and control the gene interactions and networks underlying complex cellular functions. However, current genome engineering technologies are limited in both the number and the type of perturbations that can be performed simultaneously. Here, we demonstrate that both Cas12a and a clustered regularly interspaced short palindromic repeat (CRISPR) array can be encoded in a single transcript by adding a stabilizer tertiary RNA structure. By leveraging this system, we illustrate constitutive, conditional, inducible, orthogonal and multiplexed genome engineering of endogenous targets using up to 25 individual CRISPR RNAs delivered on a single plasmid. Our method provides a powerful platform to investigate and orchestrate the sophisticated genetic programs underlying complex cell behaviors.

Original languageEnglish
Pages (from-to)887-893
Number of pages7
JournalNature Methods
Volume16
Issue number9
Publication statusPublished - Sep-2019

    Keywords

  • RNA-GUIDED ENDONUCLEASE, GENE ACTIVATION, CPF1, KNOCKOUT

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