Adenoviral vectors for in vivo delivery of CRISPR-Cas gene editors

Boucher, Paul D.; Cui, Xiaoxia; Curiel, David T.

doi:10.1016/j.jconrel.2020.09.003

Cited by 37 publications

(36 citation statements)

References 108 publications

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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].…”

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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“…In recent years, viruses have been represented as an essential and powerful tool for CRISPR due to their efficient gene delivery and long-term stable transgenic expression (Heckl et al, 2014). The most commonly utilized viral vectors are derived from adenoassociated virus (AAV) (Jarrett et al, 2018), lentivirus (LV) (Lee et al, 2021), adenovirus (Boucher et al, 2020), and baculovirus (Yin et al, 2021). These viral vectors have been widely used to deliver CRISPR/Cas9 elements for remedying genetic defects, like hearing loss (Omichi et al, 2019), neurological disorders (Pena et al, 2020), muscular dystrophies (Crudele and Chamberlain, 2019), and cystic fibrosis lung disease (Wold and Toth, 2013;Hart and Harrison, 2017).…”

Section: Viral Vectorsmentioning

confidence: 99%

“…According to their biological characteristics, they can be classified as either bioactive or abiotic. In bioactive systems, common CRISPR delivery systems contain viral vectors(Jarrett et al, 2018;Boucher et al, 2020;Lee et al, 2021), extracellular vehicles(Yao et al, 2021), cell-penetrating peptides (CPPs)(Ramakrishna et al, 2014b), or lipid nanoparticles(Cheng et al, 2020). In abiotic systems, gold nanomaterials(Wang et al, 2018), polymers(Lv et al, 2018), and graphene oxide(Yue et al, 2018) had a better effect on CRISPR system delivery.…”

mentioning

confidence: 99%

Strategies for High-Efficiency Mutation Using the CRISPR/Cas System

Feng

Wang

et al. 2022

Front. Cell Dev. Biol.

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Clustered regularly interspaced short palindromic repeats (CRISPR)-associated systems have revolutionized traditional gene-editing tools and are a significant tool for ameliorating gene defects. Characterized by high target specificity, extraordinary efficiency, and cost-effectiveness, CRISPR/Cas systems have displayed tremendous potential for genetic manipulation in almost any organism and cell type. Despite their numerous advantages, however, CRISPR/Cas systems have some inherent limitations, such as off-target effects, unsatisfactory efficiency of delivery, and unwanted adverse effects, thereby resulting in a desire to explore approaches to address these issues. Strategies for improving the efficiency of CRISPR/Cas-induced mutations, such as reducing off-target effects, improving the design and modification of sgRNA, optimizing the editing time and the temperature, choice of delivery system, and enrichment of sgRNA, are comprehensively described in this review. Additionally, several newly emerging approaches, including the use of Cas variants, anti-CRISPR proteins, and mutant enrichment, are discussed in detail. Furthermore, the authors provide a deep analysis of the current challenges in the utilization of CRISPR/Cas systems and the future applications of CRISPR/Cas systems in various scenarios. This review not only serves as a reference for improving the maturity of CRISPR/Cas systems but also supplies practical guidance for expanding the applicability of this technology.

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“…Human monocyte-derived dendritic cells (DCs), which cannot be efficiently transfected using standard transfection reagents or rapidly die after electroporation with DNA, can be transduced with HAdV-5 vectors in vitro to e.g., analyze cell type-specific functions or for genome editing using CRISPR/Cas9 [14,15]. Other approaches aim to modify the function of human DCs for the development of new vaccines by overexpressing therapeutic transgenes.…”

Section: Introductionmentioning

confidence: 99%

Breaking Entry-and Species Barriers: LentiBOOST® Plus Polybrene Enhances Transduction Efficacy of Dendritic Cells and Monocytes by Adenovirus 5

Strack

Deinzer

Thirion³

et al. 2022

Viruses

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Due to their ability to trigger strong immune responses, adenoviruses (HAdVs) in general and the serotype5 (HAdV-5) in particular are amongst the most popular viral vectors in research and clinical application. However, efficient transduction using HAdV-5 is predominantly achieved in coxsackie and adenovirus receptor (CAR)-positive cells. In the present study, we used the transduction enhancer LentiBOOST® comprising the polycationic Polybrene to overcome these limitations. Using LentiBOOST®/Polybrene, we yielded transduction rates higher than 50% in murine bone marrow-derived dendritic cells (BMDCs), while maintaining their cytokine expression profile and their capability to induce T-cell proliferation. In human dendritic cells (DCs), we increased the transduction rate from 22% in immature (i)DCs or 43% in mature (m)DCs to more than 80%, without inducing cytotoxicity. While expression of specific maturation markers was slightly upregulated using LentiBOOST®/Polybrene on iDCs, no effect on mDC phenotype or function was observed. Moreover, we achieved efficient HAdV5 transduction also in human monocytes and were able to subsequently differentiate them into proper iDCs and functional mDCs. In summary, we introduce LentiBOOST® comprising Polybrene as a highly potent adenoviral transduction agent for new in-vitro applications in a set of different immune cells in both mice and humans.

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Adenoviral vectors for in vivo delivery of CRISPR-Cas gene editors

Cited by 37 publications

References 108 publications

Boundary convection during velocity sedimentation in the Optima analytical ultracentrifuge

Boundary convection during velocity sedimentation in the Optima analytical ultracentrifuge

Strategies for High-Efficiency Mutation Using the CRISPR/Cas System

Breaking Entry-and Species Barriers: LentiBOOST® Plus Polybrene Enhances Transduction Efficacy of Dendritic Cells and Monocytes by Adenovirus 5

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