CRISPR-induced deletion with SaCas9 restores dystrophin expression in dystrophic models <i>in vitro</i> and <i>in vivo</i>

Duchêne, Benjamin; Cherif, Khadija; Iyombe-Engembe, Jean-Paul; Guyon, Antoine; Rousseau, Joël; Ouellet, Dominique L.; Barbeau, Xavier; Lagüe, Patrick; Tremblay, Jacques P.

doi:10.1101/378331

Cited by 4 publications

(5 citation statements)

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“…This line was used to test CRISPR/Cas9 genome editing complexes for reframing in exons 51 and 53 during lentiviral delivery [90]. AAV9 double SaCas9 (Staphylococcus aureus Cas9 ortholog) and guide mix was tested for deletion with borders within exons 47 and 58 for himeric exon formation [91]. Later, another mouse model with a deletion of exon 52, ∆52, was obtained using CRISPR/Cas9 genome editing system [92].…”

Section: Animal Models To Test Precision Medicine Approachesmentioning

confidence: 99%

Duchenne Muscular Dystrophy Animal Models

Егорова¹,

Galkin²,

Ivanova³

et al. 2022

Preclinical Animal Modeling in Medicine

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Duchenne muscular dystrophy is a complex and severe orphan disease. It develops when the organism lacks the expression of dystrophin - a large structural protein. Dystrophin is transcribed from the largest gene in the human genome. At the moment, there is no cure available. Dozens of groups all over the world search for cure. Animal models are an important component of both the fundamental research and therapy development. Many animal models reproducing the features of disease were created and actively used since the late 80’s until present. The species diversity spans from invertebrates to primates and the genetic diversity of these models spans from single mutations to full gene deletions. The models are often non-interchangeable; while one model may be used for particular drug design it may be useless for another. Here we describe existing models, discuss their advantages and disadvantages and potential applications for research and therapy development.

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Section: Animal Models To Test Precision Medicine Approachesmentioning

confidence: 99%

Duchenne Muscular Dystrophy Animal Models

Егорова¹,

Galkin²,

Ivanova³

et al. 2022

Preclinical Animal Modeling in Medicine

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“…To test the therapeutic potential of single-cut myoediting in vivo, we have generated mice harboring each of the most commonly deleted exons in DMD patients (35,40,41). To deliver SpCas9 and sgRNAs in vivo, we and others have used AAVs of serotype 9 (AAV9) (34,35,37,(40)(41)(42)(43)(44)(45)(46)(47)(48)(49)(50)(51). The SpCas9 cDNA is at the size limit that can be packaged into AAV9, necessitating the delivery of sgRNAs in a separate AAV9 cassette.…”

Section: Dystrophin Restoration and Correction Of Dmd In Vivomentioning

confidence: 99%

“…The SpCas9 cDNA is at the size limit that can be packaged into AAV9, necessitating the delivery of sgRNAs in a separate AAV9 cassette. Staphylococcus aureus Cas9 (SaCas9) is smaller than SpCas9 and has also been used for DMD gene editing (50,51). However, the PAM sequence for SaCas9 is more complex than that of SpCas9, which limits the potential editing events that can be introduced.…”

Section: Dystrophin Restoration and Correction Of Dmd In Vivomentioning

confidence: 99%

Toward the correction of muscular dystrophy by gene editing

Olson

2021

Proc. Natl. Acad. Sci. U.S.A.

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Recent advances in gene editing technologies are enabling the potential correction of devastating monogenic disorders through elimination of underlying genetic mutations. Duchenne muscular dystrophy (DMD) is an especially severe genetic disorder caused by mutations in the gene encoding dystrophin, a membrane-associated protein required for maintenance of muscle structure and function. Patients with DMD succumb to loss of mobility early in life, culminating in premature death from cardiac and respiratory failure. The disease has thus far defied all curative strategies. CRISPR gene editing has provided new opportunities to ameliorate the disease by eliminating DMD mutations and thereby restore dystrophin expression throughout skeletal and cardiac muscle. Proof-of-concept studies in rodents, large mammals, and human cells have validated the potential of this approach, but numerous challenges remain to be addressed, including optimization of gene editing, delivery of gene editing components throughout the musculature, and mitigation of possible immune responses. This paper provides an overview of recent work from our laboratory and others toward the genetic correction of DMD and considers the opportunities and challenges in the path to clinical translation. Lessons learned from these studies will undoubtedly enable further applications of gene editing to numerous other diseases of muscle and other tissues.

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“…This approach has great potential to develop tissue-based therapies (Rath et al, 2015). Using CRISPR-Cas9, the mutation in the dystrophin protein responsible for the most common form of Duchenne muscular dystrophy was successfully removed (Amoasii et al, 2018;Duchêne et al, 2018;Koo et al, 2018;Long et al, 2018). There are studies to prevent and treat AIDS by inhibiting the entry of HIV into the cell or by removing the HIV genome integrated into the host genome using CRISPR-Cas9 (Saayman et al, 2015).…”

Section: Treatment Of Diseasesmentioning

confidence: 99%

Bioethical issues in genome editing by CRISPR-Cas9 technology

Ayanoğlu¹,

Elçin²,

Elçin³

2020

Turk J Biol

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Biology and function of CRISPR-Cas9 technologyGenome editing technologies often work by creating fractures in chromosomal DNA. ZFN, TALEN, and CRISPR-Cas9 are all based solely on nucleases (Kim and Kim, 2014;Roh et al., 2018). The strength of these technologies stems from the ability to create fractures in the desired region of a specific target sequence as determined by the researcher. This allows researchers to modify the genome in practice in any region (Memi et al., 2018).The creation of changes in the genome depends mainly on the DNA repair capacity of the cells (Lau et al., 2018). All cells have two basic mechanisms for the repair of doubleAbstract: Genome editing technologies have led to fundamental changes in genetic science. Among them, CRISPR-Cas9 technology particularly stands out due to its advantages such as easy handling, high accuracy, and low cost. It has made a quick introduction in fields related to humans, animals, and the environment, while raising difficult questions, applications, concerns, and bioethical issues to be discussed. Most concerns stem from the use of CRISPR-Cas9 to genetically alter human germline cells and embryos (called germline genome editing). Germline genome editing leads to serial bioethical issues, such as the occurrence of undesirable changes in the genome, from whom and how informed consent is obtained, and the breeding of the human species (eugenics). However, the bioethical issues that CRISPR-Cas9 technology could cause in the environment, agriculture and livestock should also not be forgotten. In order for CRISPR-Cas9 to be used safely in all areas and to solve potential issues, worldwide legislation should be prepared, taking into account the opinions of both life and social scientists, policy makers, and all other stakeholders of the sectors, and CRISPR-Cas9 applications should be implemented according to such legislations. However, these controls should not restrict scientific freedom. Here, various applications of CRISPR-Cas9 technology, especially in medicine and agriculture, are described and ethical issues related to genome editing using CRISPR-Cas9 technology are discussed. The social and bioethical concerns in relation to human beings, other organisms, and the environment are addressed.

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CRISPR-induced deletion with SaCas9 restores dystrophin expression in dystrophic models in vitro and in vivo

Cited by 4 publications

References 64 publications

Duchenne Muscular Dystrophy Animal Models

Duchenne Muscular Dystrophy Animal Models

Toward the correction of muscular dystrophy by gene editing

Bioethical issues in genome editing by CRISPR-Cas9 technology

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