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Nat Biotechnol. 2015 Dec;33(12):1250-1255. doi: 10.1038/nbt.3412. Epub 2015 Nov 16.

Safeguarding CRISPR-Cas9 gene drives in yeast.

DiCarlo JE1,2,3, Chavez A1,2,4,5, Dietz SL1,2,4,6, Esvelt KM2,4, Church GM1,2,4.

Author information

1
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
2
Harvard Medical School, Boston, Massachusetts, USA.
3
Department of Biomedical Engineering, Boston University, Boston, Massachusetts, USA.
4
Wyss Institute for Biologically Inspired Engineering, Boston, Massachusetts, USA.
5
Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, USA.
6
Department for Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.

Abstract

RNA-guided gene drives capable of spreading genomic alterations made in laboratory organisms through wild populations could be used to address environmental and public health problems. However, the possibility of unintended genome editing occurring through the escape of strains from laboratories, coupled with the prospect of unanticipated ecological change, demands caution. We report the efficacy of CRISPR-Cas9 gene drive systems in wild and laboratory strains of the yeast Saccharomyces cerevisiae. Furthermore, we address concerns surrounding accidental genome editing by developing and validating methods of molecular confinement that minimize the risk of unwanted genome editing. We also present a drive system capable of overwriting the changes introduced by an earlier gene drive. These molecular safeguards should enable the development of safe CRISPR gene drives for diverse organisms.

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