Off-target effects in CRISPR/Cas9 gene editing

Frontiers in Bioengineering and Biotechnology, Journal Year: 2023, Volume and Issue: 11

Published: March 9, 2023

Gene editing stands for the methods to precisely make changes a specific nucleic acid sequence. With recent development of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, gene has become efficient, convenient and programmable, leading promising translational studies clinical trials both genetic non-genetic diseases. A major concern in applications CRISPR/Cas9 system is about its off-target effects, namely deposition unexpected, unwanted, or even adverse alterations genome. To date, many have been developed nominate detect sites CRISPR/Cas9, which laid basis successful upgrades derivatives with enhanced precision. In this review, we summarize these technological advancements discuss current challenges management effects future therapy.

Language: Английский

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Microfluidic technologies and devices for lipid nanoparticle-based RNA delivery

Journal of Controlled Release, Journal Year: 2022, Volume and Issue: 344, P. 80 - 96

Published: Feb. 17, 2022

Language: Английский

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Delivery of CRISPR-Cas tools for in vivo genome editing therapy: Trends and challenges

Journal of Controlled Release, Journal Year: 2022, Volume and Issue: 342, P. 345 - 361

Published: Jan. 10, 2022

The discovery of clustered regularly interspaced short palindromic repeats (CRISPR) genome editing technology opened the door to provide a versatile approach for treating multiple diseases. Promising results have been shown in numerous pre-clinical studies and clinical trials. However, safe effective method deliver genome-editing components is still key challenge vivo therapy. Adeno-associated virus (AAV) one most commonly used vector systems date, but immunogenicity against capsid, liver toxicity at high dose, potential genotoxicity caused by off-target mutagenesis genomic integration remain unsolved. Recently developed transient delivery systems, such as virus-like particle (VLP) lipid nanoparticle (LNP), may solve some issues. This review summarizes existing possible solutions overcome their limitations. Also, we highlight ongoing trials therapy recently tools applications.

Language: Английский

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In vivo delivery of CRISPR-Cas9 therapeutics: Progress and challenges

Acta Pharmaceutica Sinica B, Journal Year: 2021, Volume and Issue: 11(8), P. 2150 - 2171

Published: May 26, 2021

Within less than a decade since its inception, CRISPR-Cas9-based genome editing has been rapidly advanced to human clinical trials in multiple disease areas. Although it is highly anticipated that this revolutionary technology will bring novel therapeutic modalities many diseases by precisely manipulating cellular DNA sequences, the low efficiency of vivo delivery must be enhanced before potential can fully realized. Here we discuss most recent progress CRISPR-Cas9 systems, highlight innovative viral and non-viral technologies, emphasize outstanding challenges, provide updated perspectives.

Language: Английский

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Combinatorial design of nanoparticles for pulmonary mRNA delivery and genome editing

Nature Biotechnology, Journal Year: 2023, Volume and Issue: 41(10), P. 1410 - 1415

Published: March 30, 2023

Language: Английский

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Lipid-Nanoparticle-Based Delivery of CRISPR/Cas9 Genome-Editing Components

Molecular Pharmaceutics, Journal Year: 2022, Volume and Issue: 19(6), P. 1669 - 1686

Published: May 20, 2022

Gene editing mediated by CRISPR/Cas9 systems is due to become a beneficial therapeutic option for treating genetic diseases and some cancers. However, there are challenges in delivering CRISPR components which necessitate sophisticated delivery safe effective genome editing. Lipid nanoparticles (LNPs) have an attractive nonviral platform CRISPR-mediated their low immunogenicity application flexibility. In this review, we provide background of gene therapy, as well LNPs applicable characteristics components. We then highlight the delivery, driven significant development new, safe, optimized LNP formulations past decade. Finally, discuss considerations using deliver future perspectives on clinical translation LNP-CRISPR

Language: Английский

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In vivo somatic cell base editing and prime editing

Molecular Therapy, Journal Year: 2021, Volume and Issue: 29(11), P. 3107 - 3124

Published: Sept. 10, 2021

Recent advances in genome editing technologies have magnified the prospect of single-dose cures for many genetic diseases. For most disorders, precise DNA correction is anticipated to best treat patients. To install desired changes with high precision, our laboratory developed base editors (BEs), which can correct four common single-base substitutions, and prime editors, any substitution, insertion, and/or deletion over a stretch dozens pairs. Compared nuclease-dependent approaches that involve double-strand breaks (DSBs) often result large percentage uncontrolled outcomes, such as mixtures insertions deletions (indels), larger deletions, chromosomal rearrangements, offer greater efficiency fewer byproducts slowly dividing or non-dividing cells, those make up cells adult animals. Both viral non-viral vivo delivery methods now been used deliver animal models, establishing serve effective agents therapeutic This review summarizes examples somatic cell (post-natal) prospects future development.

Language: Английский

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Efficient in vivo base editing via single adeno-associated viruses with size-optimized genomes encoding compact adenine base editors

Nature Biomedical Engineering, Journal Year: 2022, Volume and Issue: 6(11), P. 1272 - 1283

Published: July 28, 2022

The viral delivery of base editors has been complicated by their size and the limited packaging capacity adeno-associated viruses (AAVs). Typically, dual-AAV approaches based on trans-splicing inteins have used. Here we show that, compared with systems, AAVs size-optimized genomes incorporating compact adenine (ABEs) enable efficient editing in mice at similar or lower doses. Single-AAV-encoded ABEs retro-orbitally injected led to efficiencies liver (66%), heart (33%) muscle (22%) tissues that were up 2.5-fold those ABE8e, a 93% knockdown (on average) human PCSK9 mouse Pcsk9 Angptl3 circulation, concomitant substantial reductions plasma cholesterol triglycerides. Moreover, three size-minimized ABE8e variants, each compatible single-AAV delivery, collectively offer compatibility protospacer-adjacent motifs for approximately 82% adenines genome. encoded within single will facilitate research therapeutic applications simplifying AAV production characterization, reducing dose required desired level editing.

Language: Английский

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Non-liver mRNA Delivery

Accounts of Chemical Research, Journal Year: 2021, Volume and Issue: 55(1), P. 13 - 23

Published: Dec. 3, 2021

ConspectusmRNA drugs can preempt infectious disease and treat Mendelian disorders, such as sickle cell anemia, muscular dystrophy, cystic fibrosis, well autoimmunity cancer. The three major therapeutic areas for which mRNA delivery is currently being explored are antigen production, including the COVID-19 vaccine, protein replacement therapy, genome engineering. It was demonstrated 30 years ago that introducing in vitro transcribed intramuscularly results detectable expression specific antigens protecting against likes of influenza Utilizing a modality, however, challenging. large anionic and, result, cannot passively diffuse across negatively charged plasma membrane. In addition, RNases present bloodstream tissues rapidly degrade mRNA, its administration induces innate immune response. consequence, lipid-, polymer-, dendrimer-, natural membrane-based drug systems have been developed to deliver target cells. Significant efforts investments made translate some these into clinic. Specifically, systemically administered lipid nanoparticles (LNPs) delivered liver, LNPs cells protect coronavirus 2019. However, clinically relevant non-liver spleen, lungs, heart, eye, central nervous system, lymphatics requires improved systems.In this Account, we provide an overview key advances led us Food Drug Administration approval Pfizer/BioNTech mRNA-based vaccine SARS-CoV-2 Emergency Use Authorization Moderna same disease, explain how developments will contribute clinical translation therapeutics targeted outside liver. We first focus on chemical modifications sequence optimization improve potency resulting greatly pharmacokinetics. After detailing what makes ideal payload, review used payload describe reduce clearance by obstacle development therapies. then consider recent examples tissues. Finally, discuss current programs, focusing COVID vaccines highlighting lessons may be applied future drugs.

Language: Английский

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Polymer‐Based mRNA Delivery Strategies for Advanced Therapies

Advanced Healthcare Materials, Journal Year: 2023, Volume and Issue: 12(15)

Published: Feb. 14, 2023

Abstract Messenger RNA (mRNA)‐based therapies offer great promise for the treatment of a variety diseases. In 2020, two FDA approvals mRNA‐based vaccines have elevated mRNA to global recognition. However, therapeutic capabilities extend far beyond against infectious They hold potential cancer vaccines, protein replacement therapies, gene editing and immunotherapies. For realizing such advanced it is crucial develop effective carrier systems. Recent advances in materials science led development promising nonviral delivery comparison other carriers like lipid nanoparticles, polymer‐based systems often receive less attention, despite their unique ability carefully tune chemical features promote protection, favorable pharmacokinetics, targeting delivery. this review, central highlighting molecular design criteria, stability, biodistribution are discussed. Finally, role ligands future analyzed.

Language: Английский

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