Comparison of CRISPR/Cas Endonucleases for in vivo Retinal Gene Editing

Li, Fan; Wing, Kristof; Wang, Jiang-Hui; Luu, Chi D; Bender, James; Chen, Jinying; Wang, Qi; Lu, Qinyi; Tran, Minh Thuan Nguyen; Young, Karen; Wong, Raymond Ching-Bong; Pébay, Alice; Cook, Anthony; Hung, Stevephen; Liu, Guei‐Sheung; Hewitt, Alex W.

doi:10.3389/fncel.2020.570917

Cited by 21 publications

(12 citation statements)

References 26 publications

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“…In control mice, Streptococcus pyogenes Cas9 (SpCas9) was the most efficient one using an AAV2-7m8 vector in retinal cells in vivo . 13 However, it is difficult to fit SpCas9 into one AAV. And a dual AAV system can be problematic when translating to the clinic.…”

Section: Gene Editing Tools Used For the Treatment Of Irdsmentioning

confidence: 99%

New Editing Tools for Gene Therapy in Inherited Retinal Dystrophies

2022

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Inherited retinal dystrophies (IRDs) are a heterogeneous group of diseases that affect more than 2 million people worldwide. Gene therapy (GT) has emerged as an exciting treatment modality with the potential to provide long-term benefit to patients. Today, gene addition is the most straightforward GT for autosomal recessive IRDs. However, there are three scenarios where this approach falls short. First, in autosomal dominant diseases caused by gain-of-function or dominant-negative mutations, the toxic mutated protein needs to be silenced. Second, a number of IRD genes exceed the limited carrying capacity of adeno-associated virus vectors. Third, there are still about 30% of patients with unknown mutations. In the first two contexts, precise editing tools, such as CRISPR-Cas9, base editors, or prime editors, are emerging as potential GT solutions for the treatment of IRDs. Here, we review gene editing tools based on CRISPR-Cas9 technology that have been used in vivo and the recent first-in-human application of CRISPR-Cas9 in an IRD.

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Section: Gene Editing Tools Used For the Treatment Of Irdsmentioning

confidence: 99%

New Editing Tools for Gene Therapy in Inherited Retinal Dystrophies

2022

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“…This is due to the smaller size of these enzymes being compatible with the ~4.4-kb packaging limit of the AAV genome. As alternative Cas nuclease, the small Class II, Type V Cas12a, which is able to recognize T-rich PAM sequences, can be used for viral delivery (Bin Moon et al, 2018 ; Jeon et al, 2018 ; Li et al, 2020 ).…”

Section: Editing Tools To Target Cns Cellsmentioning

confidence: 99%

Delivery Platforms for CRISPR/Cas9 Genome Editing of Glial Cells in the Central Nervous System

et al. 2021

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Glial cells (astrocytes, oligodendrocytes, and microglia) are emerging as key players in several physiological and pathological processes of the central nervous system (CNS). Astrocytes and oligodendrocytes are not only supportive cells that release trophic factors or regulate energy metabolism, but they also actively modulate critical neuronal processes and functions in the tripartite synapse. Microglia are defined as CNS-resident cells that provide immune surveillance; however, they also actively contribute to shaping the neuronal microenvironment by scavenging cell debris or regulating synaptogenesis and pruning. Given the many interconnected processes coordinated by glial cells, it is not surprising that both acute and chronic CNS insults not only cause neuronal damage but also trigger complex multifaceted responses, including neuroinflammation, which can critically contribute to the disease progression and worsening of symptoms in several neurodegenerative diseases. Overall, this makes glial cells excellent candidates for targeted therapies to treat CNS disorders. In recent years, the application of gene editing technologies has redefined therapeutic strategies to treat genetic and age-related neurological diseases. In this review, we discuss the advantages and limitations of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-based gene editing in the treatment of neurodegenerative disorders, focusing on the development of viral- and nanoparticle-based delivery methods for in vivo glial cell targeting.

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“…37,48 With the expanding repertoire of CRISPR-Cas endonucleases (SpCas9, SaCas9, Cas12a, CjCas9, and so on), the most efficacious system for target editing can be selected and optimized. 163 Compared to the expression of Cas9 from plasmid-based system or from the Cas9 mRNA, direct delivery of the synthetic Cas9 ribonucleoprotein (RNP) complex offer several advantages including its transient activity and rapid degradation with reduced off target effects, 164 in addition to the high efficiency and rapid cleavage, without the need for promoter or codon optimization. 165,166 CRISPR-Cas9 editing involves a two-step process of cleavage of the DNA followed by the DNA repair.…”

Section: Efficiencymentioning

confidence: 99%

Application of CRISPR-Cas9 Editing for Virus Engineering and the Development of Recombinant Viral Vaccines

et al. 2021

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CRISPR-Cas technology, discovered originally as a bacterial defense system, has been extensively repurposed as a powerful tool for genome editing for multiple applications in biology. In the field of virology, CRISPR-Cas9 technology has been widely applied on genetic recombination and engineering of genomes of various viruses to ask some fundamental questions of virus-host interactions. Its high efficiency, specificity, versatility, and low cost have also provided great inspirations and hope in the field of vaccinology to solve a series of bottleneck problems in the development of recombinant viral vaccines. This review highlights the applications of CRISPR editing in the technological advances compared to the traditional approaches used for the construction of recombinant viral vaccines and vectors, the main factors affecting their application, and the challenges that need to be overcome for further streamlining their effective usage in the prevention and control of diseases. Factors affecting efficiency, target specificity and fidelity of CRISPR-Cas editing in the context of viral genome editing and development of recombinant vaccines are also discussed.

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Comparison of CRISPR/Cas Endonucleases for in vivo Retinal Gene Editing

Cited by 21 publications

References 26 publications

New Editing Tools for Gene Therapy in Inherited Retinal Dystrophies

New Editing Tools for Gene Therapy in Inherited Retinal Dystrophies

Delivery Platforms for CRISPR/Cas9 Genome Editing of Glial Cells in the Central Nervous System

Application of CRISPR-Cas9 Editing for Virus Engineering and the Development of Recombinant Viral Vaccines

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