• Ming Xu, Hongzhi Xu, Jian Chen, Chunjui Chen, Feng Xu, and Zhiyong Qin.
• Department of Anesthesiology, Huashan Hospital, Fudan University, Shanghai 200040, China.
• Mol. Cell. Probes. 2018 Feb 1; 37: 32-38.

ObjectivesThis study aimed to generate mutant mice containing the Acvrl1 gene flanked with LoxP sequences to allow conditional deletion of Acvrl1 by the LoxP/Cre system. Such mice may facilitate the development of brain arteriovenous malformation (BAVM) models.MethodsThe CRISPR/Cas9 technique was used to edit Acvrl1. Two single guide RNAs (sgRNAs) with recognition sites on intron 3 and 8 and a donor vector that was homologous with the targeted gene and contained two LoxP sequences were designed and constructed. The in vitro-synthesized sgRNA, Cas9 mRNA and donor vectors were injected into mouse zygotes, which were then transferred into pseudopregnant mice. Neonatal mutant mice were identified by genotyping and sequencing.ResultsTwo mice with a floxed Acvrl1 allele were generated at a success rate of 8.7%. The target mice, which were healthy and fertile, were obtained through interbreeding.ConclusionCRISPR/Cas9 is a reliable gene-editing tool, and is able to efficiently modify Acvrl1 and create the target mice.Copyright © 2017 Elsevier Ltd. All rights reserved.

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