Efficient Generation of Mouse Models of Human Diseases Via ABE- and BE-mediated Base Editing

Overview

Journal Nat Commun

Specialty Biology

Date 2018 Jun 16

PMID 29904106

Citations 74

Authors

Zhen Liu

Zongyang Lu

Guang Yang

Shisheng Huang

Guanglei Li

Songjie Feng

Yajing Liu

Jianan Li

Wenxia Yu

Yu Zhang

Jia Chen

Qiang Sun

Xingxu Huang

Affiliations

Soon will be listed here.

Abstract

A recently developed adenine base editor (ABE) efficiently converts A to G and is potentially useful for clinical applications. However, its precision and efficiency in vivo remains to be addressed. Here we achieve A-to-G conversion in vivo at frequencies up to 100% by microinjection of ABE mRNA together with sgRNAs. We then generate mouse models harboring clinically relevant mutations at Ar and Hoxd13, which recapitulates respective clinical defects. Furthermore, we achieve both C-to-T and A-to-G base editing by using a combination of ABE and SaBE3, thus creating mouse model harboring multiple mutations. We also demonstrate the specificity of ABE by deep sequencing and whole-genome sequencing (WGS). Taken together, ABE is highly efficient and precise in vivo, making it feasible to model and potentially cure relevant genetic diseases.

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References

Lai Z, Markovets A, Ahdesmaki M, Chapman B, Hofmann O, McEwen R . VarDict: a novel and versatile variant caller for next-generation sequencing in cancer research. Nucleic Acids Res. 2016; 44(11):e108. PMC: 4914105. DOI: 10.1093/nar/gkw227. View

Kim D, Lim K, Kim S, Yoon S, Kim K, Ryu S . Genome-wide target specificities of CRISPR RNA-guided programmable deaminases. Nat Biotechnol. 2017; 35(5):475-480. DOI: 10.1038/nbt.3852. View

Kim K, Ryu S, Kim S, Baek G, Kim D, Lim K . Highly efficient RNA-guided base editing in mouse embryos. Nat Biotechnol. 2017; 35(5):435-437. DOI: 10.1038/nbt.3816. View

Li H . A statistical framework for SNP calling, mutation discovery, association mapping and population genetical parameter estimation from sequencing data. Bioinformatics. 2011; 27(21):2987-93. PMC: 3198575. DOI: 10.1093/bioinformatics/btr509. View

Dai L, Liu D, Song M, Xu X, Xiong G, Yang K . Mutations in the homeodomain of HOXD13 cause syndactyly type 1-c in two Chinese families. PLoS One. 2014; 9(5):e96192. PMC: 4006867. DOI: 10.1371/journal.pone.0096192. View

Thorvaldsdottir H, Robinson J, Mesirov J . Integrative Genomics Viewer (IGV): high-performance genomics data visualization and exploration. Brief Bioinform. 2012; 14(2):178-92. PMC: 3603213. DOI: 10.1093/bib/bbs017. View

Han G, Buchanan G, Ittmann M, Harris J, Yu X, DeMayo F . Mutation of the androgen receptor causes oncogenic transformation of the prostate. Proc Natl Acad Sci U S A. 2005; 102(4):1151-6. PMC: 544619. DOI: 10.1073/pnas.0408925102. View

Zuo E, Cai Y, Li K, Wei Y, Wang B, Sun Y . One-step generation of complete gene knockout mice and monkeys by CRISPR/Cas9-mediated gene editing with multiple sgRNAs. Cell Res. 2017; 27(7):933-945. PMC: 5518993. DOI: 10.1038/cr.2017.81. View

Zong Y, Wang Y, Li C, Zhang R, Chen K, Ran Y . Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat Biotechnol. 2017; 35(5):438-440. DOI: 10.1038/nbt.3811. View

10.

Tan M, Li J, Xu H, Melcher K, Yong E . Androgen receptor: structure, role in prostate cancer and drug discovery. Acta Pharmacol Sin. 2014; 36(1):3-23. PMC: 4571323. DOI: 10.1038/aps.2014.18. View

11.

Kim Y, Komor A, Levy J, Packer M, Zhao K, Liu D . Increasing the genome-targeting scope and precision of base editing with engineered Cas9-cytidine deaminase fusions. Nat Biotechnol. 2017; 35(4):371-376. PMC: 5388574. DOI: 10.1038/nbt.3803. View

12.

Ibrahim D, Hansen P, Rodelsperger C, Stiege A, Doelken S, Horn D . Distinct global shifts in genomic binding profiles of limb malformation-associated HOXD13 mutations. Genome Res. 2013; 23(12):2091-102. PMC: 3847778. DOI: 10.1101/gr.157610.113. View

13.

Zhao X, Sun M, Zhao J, Leyva J, Zhu H, Yang W . Mutations in HOXD13 underlie syndactyly type V and a novel brachydactyly-syndactyly syndrome. Am J Hum Genet. 2007; 80(2):361-71. PMC: 1785357. DOI: 10.1086/511387. View

14.

Gaudelli N, Komor A, Rees H, Packer M, Badran A, Bryson D . Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature. 2017; 551(7681):464-471. PMC: 5726555. DOI: 10.1038/nature24644. View

15.

Landrum M, Lee J, Benson M, Brown G, Chao C, Chitipiralla S . ClinVar: public archive of interpretations of clinically relevant variants. Nucleic Acids Res. 2015; 44(D1):D862-8. PMC: 4702865. DOI: 10.1093/nar/gkv1222. View

16.

Bae S, Park J, Kim J . Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases. Bioinformatics. 2014; 30(10):1473-5. PMC: 4016707. DOI: 10.1093/bioinformatics/btu048. View

17.

Radmayr C, Culig Z, GLATZL J, Bartsch G, Klocker H . Androgen receptor point mutations as the underlying molecular defect in 2 patients with androgen insensitivity syndrome. J Urol. 1997; 158(4):1553-6. View

18.

Flanagan S, Clauin S, Bellanne-Chantelot C, de Lonlay P, Harries L, Gloyn A . Update of mutations in the genes encoding the pancreatic beta-cell K(ATP) channel subunits Kir6.2 (KCNJ11) and sulfonylurea receptor 1 (ABCC8) in diabetes mellitus and hyperinsulinism. Hum Mutat. 2008; 30(2):170-80. DOI: 10.1002/humu.20838. View

19.

Yen S, Zhang M, Deng J, Usman S, Smith C, Parker-Thornburg J . Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes. Dev Biol. 2014; 393(1):3-9. PMC: 4166609. DOI: 10.1016/j.ydbio.2014.06.017. View

20.

Komor A, Kim Y, Packer M, Zuris J, Liu D . Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature. 2016; 533(7603):420-4. PMC: 4873371. DOI: 10.1038/nature17946. View