Heritable Multiplex Genetic Engineering in Rats Using CRISPR/Cas9

The CRISPR/Cas9 system has been proven to be an efficient gene-editing tool for genome modification of cells and organisms. Multiplex genetic engineering in rat holds a bright future for the study of complex disease. Here, we show that this system enables the simultaneous disruption of four genes (A...

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Detalles Bibliográficos
Autores principales: Ma, Yuanwu, Shen, Bin, Zhang, Xu, Lu, Yingdong, Chen, Wei, Ma, Jing, Huang, Xingxu, Zhang, Lianfeng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3943732/
https://www.ncbi.nlm.nih.gov/pubmed/24598943
http://dx.doi.org/10.1371/journal.pone.0089413
Descripción
Sumario:The CRISPR/Cas9 system has been proven to be an efficient gene-editing tool for genome modification of cells and organisms. Multiplex genetic engineering in rat holds a bright future for the study of complex disease. Here, we show that this system enables the simultaneous disruption of four genes (ApoE, B2m, Prf1, and Prkdc) in rats in one-step, by co-injection of Cas9 mRNA and sgRNAs into fertilized eggs. We further observed the gene modifications are germline transmittable, and confirmed the off-target mutagenesis and mosaicism are rarely detected by comprehensive analysis. Thus, the CRISPR/Cas9 system makes it possible to efficiently and reliably generate gene knock-out rats.

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