Titel (eng)

Fitness effects of CRISPR endonucleases in Drosophila melanogaster populations

Autor*in

Anna M. Langmüller   University of Veterinary Medicine Vienna / Cornell University

Philipp W Messer   Cornell University

Andrew G. Clark   Cornell University

Jie Du   Peking University

Yineng Xu   Cornell University

Jingxian Liu   Cornell University

Samuel E. Champer   Cornell University

Matthew Metzloff   Cornell University

Lin Xie   Cornell University

Sandra Lapinska   Cornell University

Jackson Champer   Cornell University / Peking University

Verlag

eLife Sciences Publications Ltd

Beschreibung (eng)

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 provides a highly efficient and flexible genome editing technology with numerous potential applications ranging from gene therapy to population control. Some proposed applications involve the integration of CRISPR/Cas9 endonucleases into an organism's genome, which raises questions about potentially harmful effects to the transgenic individuals. One example for which this is particularly relevant are CRISPR-based gene drives conceived for the genetic alteration of entire populations. The performance of such drives can strongly depend on fitness costs experienced by drive carriers, yet relatively little is known about the magnitude and causes of these costs. Here, we assess the fitness effects of genomic CRISPR/Cas9 expression in Drosophila melanogaster cage populations by tracking allele frequencies of four different transgenic constructs that allow us to disentangle 'direct' fitness costs due to the integration, expression, and target-site activity of Cas9, from fitness costs due to potential off-target cleavage. Using a maximum likelihood framework, we find that a model with no direct fitness costs but moderate costs due to off-target effects fits our cage data best. Consistent with this, we do not observe fitness costs for a construct with Cas9HF1, a high-fidelity version of Cas9. We further demonstrate that using Cas9HF1 instead of standard Cas9 in a homing drive achieves similar drive conversion efficiency. These results suggest that gene drives should be designed with high-fidelity endonucleases and may have implications for other applications that involve genomic integration of CRISPR endonucleases.

Sprache des Objekts

Englisch

Datum

2022

Rechte

Dieses Werk bzw. dieser Inhalt steht unter einer
CC BY 4.0 - Creative Commons Namensnennung 4.0 International Lizenz.

CC BY 4.0 International

http://creativecommons.org/licenses/by/4.0/

Klassifikation

Gene Drive; Vector; Resistance; Nucleases; Evolution; Platform; Tools; Cas9

Mitglied in der/den Collection(s) (1)