Efficient and Fast Generation of Relevant Disease Mouse Models by In Vitro and In Vivo Gene Editing of Zygotes.

Publisher: Mary Ann Liebert, Inc Country of Publication: United States NLM ID: 101738191 Publication Model: Print Cited Medium: Internet ISSN: 2573-1602 (Electronic) Linking ISSN: 25731599 NLM ISO Abbreviation: CRISPR J Subsets: MEDLINE

Original Publication: [New Rochelle, NY] : Mary Ann Liebert, Inc., [2018]-

Gene Editing*/methods ; Zygote*/metabolism; Animals ; CRISPR-Cas Systems/genetics ; Mice ; Mice, Knockout ; Ribonucleoproteins/genetics ; RNA, Guide, CRISPR-Cas Systems

Knockout mice for human disease-causing genes provide valuable models in which new therapeutic approaches can be tested. Electroporation of genome editing tools into zygotes, in vitro or within oviducts, allows for the generation of targeted mutations in a shorter time. We have generated mouse models deficient in genes involved in metabolic rare diseases (Primary Hyperoxaluria Type 1 Pyruvate Kinase Deficiency) or in a tumor suppressor gene ( Rasa1 ). Pairs of guide RNAs were designed to generate controlled deletions that led to the absence of protein. In vitro or in vivo ribonucleoprotein (RNP) electroporation rendered more than 90% and 30% edited newborn animals, respectively. Mice lines with edited alleles were established and disease hallmarks have been verified in the three models that showed a high consistency of results and validating RNP electroporation into zygotes as an efficient technique for disease modeling without the need to outsource to external facilities.

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0 (Ribonucleoproteins)

Date Created: 20220610 Date Completed: 20220614 Latest Revision: 20240221

20240221

PMC9233508

10.1089/crispr.2022.0013

35686982