Translating CRISPR-Cas Therapeutics: Approaches and Challenges

Tay, Lavina Sierra; Palmer, Nathan; Panwala, Rebecca; Chew, Wei Leong; Mali, Prashant

doi:10.1089/crispr.2020.0025

Cited by 27 publications

(30 citation statements)

References 273 publications

(264 reference statements)

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“…Recent developments in gene therapy over the last decade or so has drawn the renewed attention of many companies and researchers to the use of viral vectors (using various virus systems, e.g., adenovirus [44][45][46][47][48] and AAV [49][50][51]), along with completely synthetic particles, involving liposomes and nanoparticles [52][53][54][55][56][57], as drug (nucleic acid) delivery systems. Initially, interest in this area was predominately concerned with the delivery of DNA therapeutic material, however, interest has widened to include the use of RNA therapeutics [58] such as interference ribonucleic acids, RNAi [59,60], and messenger ribonucleic acids, mRNA [61,62] along with gene editing payload material [63][64][65][66]. Given the success and potential offered by these new classes of biopharmaceuticals, they are revolutionizing drug development.…”

Section: Discussionmentioning

confidence: 99%

Boundary convection during velocity sedimentation in the Optima analytical ultracentrifuge

Berkowitz¹,

Laue

2021

Preprint

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Analytical ultracentrifugation (AUC) provides the most widely applicable, precise and accurate means for characterizing solution hydrodynamic and thermodynamic properties. In recent times AUC has found broad application in the biopharmaceutical industry as a first-principle means for quantitatively characterizing biopharmaceuticals. Boundary sedimentation velocity AUC (SV-AUC) analysis is widely used to assess protein aggregation, fragmentation and conformational variants in the same solvents used during drug development and production. SV-AUC is especially useful for the analysis of drug substance, drug product and dosing solution, where other techniques may exhibit solvent matrix issues or concentration limitations. Recently, the only manufacturer of the analytical ultracentrifuge, released its newest (third generation) analytical ultracentrifuge, the Optima, in early 2017 to replace its aging 2nd generation XL series ultracentrifuges. However, SV-AUC data from four Optima units used in conducting characterization work on adeno-associated virus (AAV) has shown evidence of sample convection. Further investigation reveals that this problem arises from the temperature control system design, which is prone to producing destabilizing temperature induced density gradients that can lead to density inversions. The observed convection impacts both the qualitative and quantitative data generated by the Optima. The problem is intermittent and variable in severity within a given Optima unit and between Optima units. This convection appears to be mainly associated with low rotor speeds and dilute samples in dilute solvents, such as AAV samples in formulation buffers containing relatively low concentrations of salts, sugars, etc. Under these conditions it is found that a sufficiently robust stabilizing density gradient is not always present during sedimentation, making the sample susceptible to convection by localized density inversions. Because SV-AUC is used as an analytical tool in making critical decisions in the development and quality control of biotherapeutics, it is imperative to alert users about this potential problem. In general special attention to data quality needs to be made by those researchers working with very large biopharmaceutical particles (e.g. gene therapy products that involve viral vectors or nanoparticles), where the conditions leading to convection are most likely to occur. It is important to note that the XL series analytical ultracentrifuges do not suffer from this problem, indicating that this problem is unique to the Optima. Attributes that reveal the presence of this problem and strategies for its elimination or minimization are provided.

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Section: Discussionmentioning

confidence: 99%

Boundary convection during velocity sedimentation in the Optima analytical ultracentrifuge

Berkowitz¹,

Laue

2021

Preprint

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“…Inorganic nanoparticles on the other hand, are easier to manufacture as compared to viral and lipid/polymer-based vectors, as they have a more predictable composition, size, and dispersion, are easier to describe and structurally flocculate, and are much more consistent over time. As a result, future investigations are directed on the in vitro and in vivo use of inorganic nanoparticles as CRISPR/Cas9 delivery vehicles [206].…”

Section: Novel Delivery Methods For Retention and Higher Efficacymentioning

confidence: 99%

“…CRISPR Gold Cas9 RNP with prototype DNA was infused intramuscularly and repaired 5.4% of dystrophin genetic mutations. In a mouse model of autism spectrum disorder (ASD), intracranial infusion of CRISPR Gold Cas9 RNP caused targeted gene editing of 14.5% of the GRM5 gene and regained 40-50% of protein synthesis in the brain and other organs, reversing the consequences of ASD [205,206].…”

Section: Metal Nanoparticles For Efficient Crispr/cas9 Deliverymentioning

confidence: 99%

Exploring nano-enabled CRISPR-Cas-powered strategies for efficient diagnostics and treatment of infectious diseases

et al. 2022

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Biomedical researchers have subsequently been inspired the development of new approaches for precisely changing an organism's genomic DNA in order to investigate customized diagnostics and therapeutics utilizing genetic engineering techniques. Clustered Regulatory Interspaced Short Palindromic Repeats (CRISPR) is one such technique that has emerged as a safe, targeted, and effective pharmaceutical treatment against a wide range of disease-causing organisms, including bacteria, fungi, parasites, and viruses, as well as genetic abnormalities. The recent discovery of very flexible engineered nucleic acid binding proteins has changed the scientific area of genome editing in a revolutionary way. Since current genetic engineering technique relies on viral vectors, issues about immunogenicity, insertional oncogenesis, retention, and targeted delivery remain unanswered. The use of nanotechnology has the potential to improve the safety and efficacy of CRISPR/ Cas9 component distribution by employing tailored polymeric nanoparticles. The combination of two (CRISPR/Cas9 and nanotechnology) offers the potential to open new therapeutic paths. Considering the benefits, demand, and constraints, the goal of this research is to acquire more about the biology of CRISPR technology, as well as aspects of selective and effective diagnostics and therapies for infectious illnesses and other metabolic disorders. This review advocated combining nanomedicine (nanomedicine) with a CRISPR/Cas enabled sensing system to perform early-stage diagnostics and selective therapy of specific infectious disorders. Such a Nano-CRISPR-powered nanomedicine and sensing system would allow for successful infectious illness control, even on a personal level. This comprehensive study also discusses the current obstacles and potential of the predicted technology.

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“…Excluding the actual limitations existing in the current approaches, another point of concern is the fact that some off-targets may be completely benign, whereas others could have serious consequences depending on the cell context or the indication. This is a well-recognized issue in the field and is currently being addressed by engineering the CRISPR payload at both the protein and gRNA level with simultaneous optimization of the ideal window of active exposure of the cells of interest to the functional RNP complex (Tay et al, 2020 ).…”

Section: The Era Of Genome Editing: Challenges and Prospectsmentioning

confidence: 99%

The Promise and the Hope of Gene Therapy

Papanikolaou

Bosio

2021

Front. Genome Ed.

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It has been over 30 years since visionary scientists came up with the term “Gene Therapy,” suggesting that for certain indications, mostly monogenic diseases, substitution of the missing or mutated gene with the normal allele via gene addition could provide long-lasting therapeutic effect to the affected patients and consequently improve their quality of life. This notion has recently become a reality for certain diseases such as hemoglobinopathies and immunodeficiencies and other monogenic diseases. However, the therapeutic wave of gene therapies was not only applied in this context but was more broadly employed to treat cancer with the advent of CAR-T cell therapies. This review will summarize the gradual advent of gene therapies from bench to bedside with a main focus on hemopoietic stem cell gene therapy and genome editing and will provide some useful insights into the future of genetic therapies and their gradual integration in the everyday clinical practice.

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Translating CRISPR-Cas Therapeutics: Approaches and Challenges

Cited by 27 publications

References 273 publications

Boundary convection during velocity sedimentation in the Optima analytical ultracentrifuge

Boundary convection during velocity sedimentation in the Optima analytical ultracentrifuge

Exploring nano-enabled CRISPR-Cas-powered strategies for efficient diagnostics and treatment of infectious diseases

The Promise and the Hope of Gene Therapy

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