High-fidelity Endonuclease Variant HypaCas9 Facilitates Accurate Allele-specific Gene Modification in Mouse Zygotes

Overview

Journal Commun Biol

Specialty Biology

Date 2019 Oct 22

PMID 31633062

Citations 27

Authors

Arisa Ikeda

Wataru Fujii

Koji Sugiura

Kunihiko Naito

Affiliations

Soon will be listed here.

Abstract

CRISPR/Cas9 has been widely used for the efficient generation of genetically modified animals; however, this system could have unexpected off-target effects. In the present study, we confirmed the validity of a high-fidelity Cas9 variant, HypaCas9, for accurate genome editing in mouse zygotes. HypaCas9 efficiently modified the target locus while minimizing off-target effects even in a single-nucleotide mismatched sequence. Furthermore, by applying HypaCas9 to the discrimination of SNP in hybrid strain-derived zygotes, we accomplished allele-specific gene modifications and successfully generated mice with a monoallelic mutation in an essential gene. These results suggest that the improved accuracy of HypaCas9 facilitates the generation of genetically modified animals.

Citing Articles

Leveraging CRISPR gene editing technology to optimize the efficacy, safety and accessibility of CAR T-cell therapy.

Lei T, Wang Y, Zhang Y, Yang Y, Cao J, Huang J Leukemia. 2024; 38(12):2517-2543.

PMID: 39455854 PMC: 11588664. DOI: 10.1038/s41375-024-02444-y.

Perspectives on CRISPR Genome Editing to Prevent Prion Diseases in High-Risk Individuals.

Medd M, Cao Q Biomedicines. 2024; 12(8).

PMID: 39200190 PMC: 11352000. DOI: 10.3390/biomedicines12081725.

Allele-specific CRISPR-Cas9 editing inactivates a single nucleotide variant associated with collagen VI muscular dystrophy.

Bolduc V, Sizov K, Brull A, Esposito E, Chen G, Uapinyoying P Mol Ther Nucleic Acids. 2024; 35(3):102269.

PMID: 39171142 PMC: 11338111. DOI: 10.1016/j.omtn.2024.102269.

Allele-specific CRISPR/Cas9 editing inactivates a single nucleotide variant associated with collagen VI muscular dystrophy.

Bolduc V, Sizov K, Brull A, Esposito E, Chen G, Uapinyoying P bioRxiv. 2024; .

PMID: 38585815 PMC: 10996683. DOI: 10.1101/2024.03.22.586265.

Current Strategies for Increasing Knock-In Efficiency in CRISPR/Cas9-Based Approaches.

Leal A, Herreno-Pachon A, Benincore-Florez E, Karunathilaka A, Tomatsu S Int J Mol Sci. 2024; 25(5).

PMID: 38473704 PMC: 10931195. DOI: 10.3390/ijms25052456.

References

Hwang W, Fu Y, Reyon D, Maeder M, Tsai S, Sander J . Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat Biotechnol. 2013; 31(3):227-9. PMC: 3686313. DOI: 10.1038/nbt.2501. View

Kleinstiver B, Pattanayak V, Prew M, Tsai S, Nguyen N, Zheng Z . High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016; 529(7587):490-5. PMC: 4851738. DOI: 10.1038/nature16526. View

Li W, Teng F, Li T, Zhou Q . Simultaneous generation and germline transmission of multiple gene mutations in rat using CRISPR-Cas systems. Nat Biotechnol. 2013; 31(8):684-6. DOI: 10.1038/nbt.2652. View

Brinkman E, Chen T, Amendola M, van Steensel B . Easy quantitative assessment of genome editing by sequence trace decomposition. Nucleic Acids Res. 2014; 42(22):e168. PMC: 4267669. DOI: 10.1093/nar/gku936. View

Yang Q, Fujii W, Kaji N, Kakuta S, Kada K, Kuwahara M . The essential role of phospho-T38 CPI-17 in the maintenance of physiological blood pressure using genetically modified mice. FASEB J. 2017; 32(4):2095-2109. DOI: 10.1096/fj.201700794R. View

Dehairs J, Talebi A, Cherifi Y, Swinnen J . CRISP-ID: decoding CRISPR mediated indels by Sanger sequencing. Sci Rep. 2016; 6:28973. PMC: 4929496. DOI: 10.1038/srep28973. View

Kulcsar P, Talas A, Huszar K, Ligeti Z, Toth E, Weinhardt N . Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage. Genome Biol. 2017; 18(1):190. PMC: 6389135. DOI: 10.1186/s13059-017-1318-8. View

Satyanarayana A, Berthet C, Lopez-Molina J, Coppola V, Tessarollo L, Kaldis P . Genetic substitution of Cdk1 by Cdk2 leads to embryonic lethality and loss of meiotic function of Cdk2. Development. 2008; 135(20):3389-400. PMC: 2668819. DOI: 10.1242/dev.024919. View

Fujii W, Onuma A, Sugiura K, Naito K . Efficient generation of genome-modified mice via offset-nicking by CRISPR/Cas system. Biochem Biophys Res Commun. 2014; 445(4):791-4. DOI: 10.1016/j.bbrc.2014.01.141. View

10.

Hara S, Tamano M, Yamashita S, Kato T, Saito T, Sakuma T . Generation of mutant mice via the CRISPR/Cas9 system using FokI-dCas9. Sci Rep. 2015; 5:11221. PMC: 4460908. DOI: 10.1038/srep11221. View

11.

Dominguez A, Lim W, Qi L . Beyond editing: repurposing CRISPR-Cas9 for precision genome regulation and interrogation. Nat Rev Mol Cell Biol. 2015; 17(1):5-15. PMC: 4922510. DOI: 10.1038/nrm.2015.2. View

12.

Wang H, Yang H, Shivalila C, Dawlaty M, Cheng A, Zhang F . One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell. 2013; 153(4):910-8. PMC: 3969854. DOI: 10.1016/j.cell.2013.04.025. View

13.

Fujii W, Ikeda A, Sugiura K, Naito K . Efficient Generation of Genome-Modified Mice Using Campylobacter jejuni-Derived CRISPR/Cas. Int J Mol Sci. 2017; 18(11). PMC: 5713256. DOI: 10.3390/ijms18112286. View

14.

Vakulskas C, Dever D, Rettig G, Turk R, Jacobi A, Collingwood M . A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells. Nat Med. 2018; 24(8):1216-1224. PMC: 6107069. DOI: 10.1038/s41591-018-0137-0. View

15.

Casini A, Olivieri M, Petris G, Montagna C, Reginato G, Maule G . A highly specific SpCas9 variant is identified by in vivo screening in yeast. Nat Biotechnol. 2018; 36(3):265-271. PMC: 6066108. DOI: 10.1038/nbt.4066. View

16.

Nishimasu H, Shi X, Ishiguro S, Gao L, Hirano S, Okazaki S . Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science. 2018; 361(6408):1259-1262. PMC: 6368452. DOI: 10.1126/science.aas9129. View

17.

Nakagawa Y, Sakuma T, Sakamoto T, Ohmuraya M, Nakagata N, Yamamoto T . Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. BMC Biotechnol. 2015; 15:33. PMC: 4440308. DOI: 10.1186/s12896-015-0144-x. View

18.

Shen B, Zhang J, Wu H, Wang J, Ma K, Li Z . Generation of gene-modified mice via Cas9/RNA-mediated gene targeting. Cell Res. 2013; 23(5):720-3. PMC: 3641603. DOI: 10.1038/cr.2013.46. View

19.

Fujii W . Generation of Knock-in Mouse by Genome Editing. Methods Mol Biol. 2017; 1630:91-100. DOI: 10.1007/978-1-4939-7128-2_8. View

20.

Fujii W, Kakuta S, Yoshioka S, Kyuwa S, Sugiura K, Naito K . Zygote-mediated generation of genome-modified mice using Streptococcus thermophilus 1-derived CRISPR/Cas system. Biochem Biophys Res Commun. 2016; 477(3):473-6. DOI: 10.1016/j.bbrc.2016.06.070. View