CRISPR/Cas9 Technology on Cardiovascular Research    

Cardiovascular disease (CVD) is the leading cause of mortality worldwide. CVD describes a group of diseases affecting the heart and blood vessels, such as coronary artery disease, cardiomyopathies, heart failure or arrhythmias. Various risk factors, such as smoking, obesity, hypertension or high cholesterol can be causative for CVD, nevertheless, it is understood that these traditional risk factors only contribute to a fraction of disease cases. However, further mechanistic studies aimed at understanding the cause of the diseases are limited by the fact that the isolation and culture of human primary cardiomyocytes for cardiovascular research is exceedingly difficult. In this case, researchers all over the world have invested time and resources to ameliorate animal and cell models for cardiac diseases.

The CRISPR/Cas9 System in Cardiovascular Diseases

Genome editing has rapidly emerged as a potent tool in basic and translational research. Zinc finger nucleases and transcription activator-like effector nucleases (TALENs) catalyzed the field initially. With the development of the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) system, the field is expanding even more rapidly due to its specificity, efficacy, and ease of use. Generally, genome editing tools induce site-specific DNA double-strand breaks at a specific genomic site, leading to the activation of the nonhomologous end-joining (NHEJ) and homologous recombination (HR) cellular endogenous double-strand break repair machinery. Recent advances of the CRISPR technology also allow for RNA recognition, making it possible to cleave RNA, enhance or inhibit translation, induce specific post-transcriptional modifications, or support isolation of specific RNA:protein complexes.

Figure 1. Genome editing in Cardiovascular Diseases

The CRISPR/Cas9 technology has been used to create mouse models of genetic diseases, such as severe cardiomyopathy, in a shorter time than with the traditional homologous recombination technique. Besides, it is now possible to inject CRISPR/Cas9 into cell embryos in rats, rabbits, and primates, which are used to study cardiovascular diseases. With the emergence of induced pluripotent stem cell (iPSC) technology, the researchers have also been able to generate alternative cell models to investigate the molecular mechanisms of the diseases. In this condition, the CRISPR/Cas9 technology provides a straightforward mechanism to explain how the cells misbehave by allowing the reversion of the causal mutation.

Therapeutic Potential of CRISPR/Cas9 Technology in Cardiovascular Diseases

The application of CRISPR/Cas9 to modulate cardiovascular cells opens up a new era of opportunity for regenerative cardiovascular medicine. Abrahimi et al. used endothelial colony forming cell derived-EC and CRISPR/Cas9 system to create EC lacking MHC class II transactivator (CIITA). These ECs cannot stimulate allogeneic CD4⁺ T cells but retain the ability to form vascular structure, thus the engineered ECs are non-immunogenic and can be used for blood vessel reconstruction. Similarly, CRISPR/Cas9 edited B2M and CCR5 in CD34⁺ hematopoietic progenitor cells retain multilineage potential and can be used for hematopoietic progenitor cell-based therapy in the future to treat a variety of conditions including ischemic heart diseases.

Duchenne muscular dystrophy (DMD) is an X-linked genetic disorder associated with both cardiomyopathy and skeletal muscle wasting. Long et al. suggested that the dystrophy gene can be corrected by CRISPR/Cas9 mediated gene editing in the germ line of MDX mice, a mouse model for DMD. Moreover, Ding et al. proved that CRISPR/Cas9 can be used to edit genes in somatic cells in vivo. They disrupt the proprotein convertase subtilisin/kexin type 9 (PCSK9) gene and reduce blood cholesterol levels only a few days after administration of an adenovirus expressing Cas9 and guide RNA. This proof of principle study to target PCSK9 in somatic cells in vivo showed that CRISPR/Cas9 system has enormous therapeutic potential in preventing CHD by a single shot and this is far more convenient than taking statin regularly.

Our CRISPR/Cas9 System Services

CRISPR/Cas9 Platform^CB, a global leading genetic editing biotechnology company, is committed to providing the most professional and comprehensive genetic editing technology solutions for our clients. To support your projects, we offer a comprehensive custom CRISPR/Cas9 gene editing service from strategy design to final cardiovascular disease model generation.

• Human Cell Models Generation of Cardiovascular Disease by CRISPR/Cas9 System

The CRISPR/Cas9 tool allows for a relatively efficient and easy generation of isogenic cell lines differing only by the DNA sequence of interest, thereby eliminating other confounders like genetic background and epigenetic memory. So far, the CRISPR/Cas9 system has already been proven to be an effective approach to create gene knockout or knockin in human cells and in particular in iPSCs. CRISPR/Cas9 Platform^CB can combine the CRISPR/Cas9 system and iPSC technology to generate better in vitro models of cardiac pathologies.

• Animal Models Generation of Cardiovascular Disease by CRISPR/Cas9 System

The development of CRISPR/Cas9 technology has represented a powerful breakthrough for the generation of diverse animal models to the aim of heart disease investigation. We offer custom animal models of cardiovascular disease by CRISPR/Cas9 system to meet different needs. We have enabled the establishment of very effective protocols that are completely reliable for the generation of knockout and knockin point mutations useful for cardiac gene editing, cardiac disease modeling, and for exploration of potential gene therapy.
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Animal Models

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Cell Lines

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