Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Homology-directed repair of an MYBPC3 gene mutation in a rat model of hypertrophic cardiomyopathy


Variants in myosin-binding protein C3 (MYBPC3) gene are a main cause of hypertrophic cardiomyopathy (HCM), accounting for 30% to 40% of the total number of HCM mutations. Gene editing represents a potential permanent cure for HCM. The aim of this study was to investigate whether genome editing of MYBPC3 using the CRISPR/Cas9 system in vivo could rescue the phenotype of rats with HCM. We generated a rat model of HCM (“1098hom”) that carried an Mybpc3 premature termination codon mutation (p.W1098x) discovered in a human HCM pedigree. On postnatal day 3, the CRISPR/Cas9 system was introduced into rat pups by a single dose of AAV9 particles to correct the variant using homology-directed repair (HDR). Analysis was performed 6 months after AAV9 injection. The 1098hom rats didn’t express MYBPC3 protein and developed an HCM phenotype with increased ventricular wall thickness and diminished cardiac function. Importantly, CRISPR HDR genome editing corrected 3.56% of total mutations, restored MYBPC3 protein expression by 2.12%, and normalized the HCM phenotype of 1098hom rats. Our work demonstrates that the HDR strategy is a promising approach for treating HCM associated with MYBPC3 mutation, and that CRISPR technology has great potential for treating hereditary heart diseases.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Get just this article for as long as you need it


Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Generation of the 1098hom rat model of HCM.
Fig. 2: Designation of CRISPR/Cas9 HDR strategy to correct the p.W1098x mutation.
Fig. 3: HDR treatment alleviated cardiac hypertrophy in 1098hom rats.
Fig. 4: HDR treatment improved cardiac function in 1098hom rats.
Fig. 5: Evaluation of HDR editing efficiency.
Fig. 6: HDR treatment partially restored MYBPC3 protein expression.

Data availability

The datasets analyzed during the current study are available from the corresponding author on reasonable request.


  1. Maron BJ, Ommen SR, Semsarian C, Spirito P, Olivotto I, Maron MS. Hypertrophic cardiomyopathy: present and future, with translation into contemporary cardiovascular medicine. J Am Coll Cardiol. 2014;64:83–99.

    Article  PubMed  Google Scholar 

  2. Marian AJ, Braunwald E. Hypertrophic cardiomyopathy: genetics, pathogenesis, clinical manifestations, diagnosis, and therapy. Circ Res. 2017;121:749–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Lopes LR, Zekavati A, Syrris P, Hubank M, Giambartolomei C, Dalageorgou C, et al. Genetic complexity in hypertrophic cardiomyopathy revealed by high-throughput sequencing. J Med Genet. 2013;50:228–39.

    Article  CAS  PubMed  Google Scholar 

  4. Girolami F, Ho CY, Semsarian C, Baldi M, Will ML, Baldini K, et al. Clinical features and outcome of hypertrophic cardiomyopathy associated with triple sarcomere protein gene mutations. J Am Coll Cardiol. 2010;55:1444–53.

    Article  CAS  PubMed  Google Scholar 

  5. Ho CY, Charron P, Richard P, Girolami F, Van Spaendonck-Zwarts KY, Pinto Y. Genetic advances in sarcomeric cardiomyopathies: state of the art. Cardiovasc Res. 2015;105:397–408.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Sarikas A, Carrier L, Schenke C, Doll D, Flavigny J, Lindenberg KS, et al. Impairment of the ubiquitin-proteasome system by truncated cardiac myosin binding protein C mutants. Cardiovasc Res. 2005;66:33–44.

    Article  CAS  PubMed  Google Scholar 

  7. Millat G, Bouvagnet P, Chevalier P, Dauphin C, Jouk PS, Da Costa A, et al. Prevalence and spectrum of mutations in a cohort of 192 unrelated patients with hypertrophic cardiomyopathy. Eur J Med Genet. 2010;53:261–7.

    Article  PubMed  Google Scholar 

  8. Gao X, Tao Y, Lamas V, Huang M, Yeh WH, Pan B, et al. Treatment of autosomal dominant hearing loss by in vivo delivery of genome editing agents. Nature. 2018;553:217–21.

    Article  CAS  PubMed  Google Scholar 

  9. Yang Y, Wang L, Bell P, McMenamin D, He Z, White J, et al. A dual AAV system enables the Cas9-mediated correction of a metabolic liver disease in newborn mice. Nat Biotechnol. 2016;34:334–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Amoasii L, Long C, Li H, Mireault AA, Shelton JM, Sanchez-Ortiz E, et al. Single-cut genome editing restores dystrophin expression in a new mouse model of muscular dystrophy. Sci Transl Med. 2017;9:eaan8081.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Noordzij M, Dekker FW, Zoccali C, Jager KJ. Sample size calculations. Nephron Clin Pract. 2011;118:c319–23.

    Article  PubMed  Google Scholar 

  12. Stemmer M, Thumberger T, Del Sol Keyer M, Wittbrodt J, Mateo JL. CCTop: an intuitive, flexible and reliable CRISPR/Cas9 target prediction tool. PloS One. 2015;10:e0124633.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Labuhn M, Adams FF, Ng M, Knoess S, Schambach A, Charpentier EM, et al. Refined sgRNA efficacy prediction improves large- and small-scale CRISPR-Cas9 applications. Nucl Acids Res. 2018;46:1375–85.

    Article  CAS  PubMed  Google Scholar 

  14. Drittanti L, Rivet C, Manceau P, Danos O, Vega M. High throughput production, screening and analysis of adeno-associated viral vectors. Gene Therapy. 2000;7:924–9.

    Article  CAS  PubMed  Google Scholar 

  15. Yardeni T, Eckhaus M, Morris HD, Huizing M, Hoogstraten-Miller S. Retro-orbital injections in mice. Lab Animal. 2011;40:155–60.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Zhao H, Li Y, He L, Pu W, Yu W, Li Y, et al. In vivo AAV-CRISPR/Cas9-mediated gene editing ameliorates atherosclerosis in familial hypercholesterolemia. Circulation. 2020;141:67–79.

    Article  CAS  PubMed  Google Scholar 

  17. Yin H, Xue W, Chen S, Bogorad RL, Benedetti E, Grompe M, et al. Genome editing with Cas9 in adult mice corrects a disease mutation and phenotype. Nat Biotechnol. 2014;32:551–3.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Cai Y, Cheng T. In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway. Sci Adv. 2019;5:eaav3335.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Lee K, Conboy M, Park HM, Jiang F, Kim HJ, Dewitt MA, et al. Nanoparticle delivery of Cas9 ribonucleoprotein and donor DNA in vivo induces homology-directed DNA repair. Nat. Biomedi Eng. 2017;1:889–901.

    Article  CAS  Google Scholar 

  20. Bergmann O, Bhardwaj RD, Bernard S, Zdunek S, Barnabe-Heider F, Walsh S, et al. Evidence for cardiomyocyte renewal in humans. Science. 2009;324:98–102.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Payan SM, Hubert F, Rochais F. Cardiomyocyte proliferation, a target for cardiac regeneration. Biochim Biophys Acta Mol Cell Res. 2020;1867:118461.

  22. Long C, Amoasii L, Mireault AA, McAnally JR, Li H, Sanchez-Ortiz E, et al. Postnatal genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy. Science. 2016;351:400–3.

    Article  CAS  PubMed  Google Scholar 

  23. van Putten M, van der Pijl EM, Hulsker M, Verhaart IE, Nadarajah VD, van der Weerd L, et al. Low dystrophin levels in heart can delay heart failure in mdx mice. J Mol Cell Cardiol. 2014;69:17–23.

    Article  PubMed  Google Scholar 

  24. Xie C, Zhang YP, Song L, Luo J, Qi W, Hu J, et al. Genome editing with CRISPR/Cas9 in postnatal mice corrects PRKAG2 cardiac syndrome. Cell Res. 2016;26:1099–111.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Carrier L, Mearini G, Stathopoulou K, Cuello F. Cardiac myosin-binding protein C (MYBPC3) in cardiac pathophysiology. Gene. 2015;573:188–97.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Prondzynski M, Mearini G, Carrier L. Gene therapy strategies in the treatment of hypertrophic cardiomyopathy. Pflugers Archiv Eur J Physiol. 2019;471:807–15.

    Article  CAS  Google Scholar 

Download references


We acknowledge and appreciate our colleagues for their valuable suggestions and technical assistance for this study.


This work was supported by National Natural Science Foundation of China [No. 82100401, 82070354, 81470519, 81630010] and Huazhong University of Science and Technology Academic Frontier Youth Team (No. 2019QYTD08).

Author information

Authors and Affiliations



DWW, LN and JN designed the study. JN and YH conducted the experiments, analyzed data and completed the manuscript. ZJ, WH and HS conducted the animal experiments. ZW performed the echocardiography examination of the animals.

Corresponding authors

Correspondence to Li Ni or Dao Wen Wang.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval

The study was approved by the Ethics Review Board of Tongji Hospital and Tongji Medical College. It complied with the principles of the Declaration of Helsinki. Written informed consent was obtained from individual subjects of the HCM pedigree in this study. All animal experiments complied with the “Guide for the Care and Use of Laboratory Animals” published by the United States National Institutes of Health (NIH Publication No. 85-23, revised 1996). This study was approved by the Institutional Animal Research Committee of Tongji Medical College.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nie, J., Han, Y., Jin, Z. et al. Homology-directed repair of an MYBPC3 gene mutation in a rat model of hypertrophic cardiomyopathy. Gene Ther (2023).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI:


Quick links