Applications of genome editing technology in the targeted therapy of human diseases: mechanisms, advances and prospects

Signal Transduction and Targeted Therapy, Journal Year: 2020, Volume and Issue: 5(1)

Published: Jan. 3, 2020

Abstract Based on engineered or bacterial nucleases, the development of genome editing technologies has opened up possibility directly targeting and modifying genomic sequences in almost all eukaryotic cells. Genome extended our ability to elucidate contribution genetics disease by promoting creation more accurate cellular animal models pathological processes begun show extraordinary potential a variety fields, ranging from basic research applied biotechnology biomedical research. Recent progress developing programmable such as zinc-finger nucleases (ZFNs), transcription activator-like effector (TALENs) clustered regularly interspaced short palindromic repeat (CRISPR)–Cas-associated greatly expedited gene concept clinical practice. Here, we review recent advances three major (ZFNs, TALENs, CRISPR/Cas9) discuss applications their derivative reagents tools various human diseases future therapies, focusing cells models. Finally, provide an overview trials applying platforms for treatment some challenges implementation this technology.

Language: Английский

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Systemic nanoparticle delivery of CRISPR-Cas9 ribonucleoproteins for effective tissue specific genome editing

Nature Communications, Journal Year: 2020, Volume and Issue: 11(1)

Published: June 26, 2020

Abstract CRISPR-Cas9 has emerged as a powerful technology that relies on Cas9/sgRNA ribonucleoprotein complexes (RNPs) to target and edit DNA. However, many therapeutic targets cannot currently be accessed due the lack of carriers can deliver RNPs systemically. Here, we report generalizable methodology allows engineering modified lipid nanoparticles efficiently into cells tissues including muscle, brain, liver, lungs. Intravenous injection facilitated tissue-specific, multiplexed editing six genes in mouse High carrier potency was leveraged create organ-specific cancer models livers lungs mice though facile knockout multiple genes. The developed were also able restore dystrophin expression DMD significantly decrease serum PCSK9 level C57BL/6 mice. Application this strategy will facilitate broad nanoparticle development for variety disease amenable protein delivery precise gene correction approaches.

Language: Английский

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Lipid Nanoparticles Enabling Gene Therapies: From Concepts to Clinical Utility

Nucleic Acid Therapeutics, Journal Year: 2018, Volume and Issue: 28(3), P. 146 - 157

Published: April 23, 2018

Genetic drugs based on RNA or DNA have remarkable therapeutic potential as virtually any disease can be treated by silencing a pathological gene, expressing beneficial protein, editing defective genes. However, therapies nucleic acid polymers require sophisticated delivery systems to deliver these macromolecules the interior of target cells. In this study, we review progress in developing nonviral lipid nanoparticle (LNP) that attractive properties, including ease manufacture, reduced immune responses, multidosing capabilities, larger payloads, and flexibility design. LNP represent most advanced for genetic it is expected an LNP-short interfering (siRNA) formulation will receive clinical approval from Food Drug Administration (FDA) 2018 treatment hereditary condition transthyretin-mediated amyloidosis, fatal which there currently no treatment. This achievement largely due development optimized ionizable cationic lipids, arguably important factor success LNP-siRNA. addition, highlight applications, targeting tissues beyond liver approaches messenger Clustered Regularly Interspaced Short Palindromic Repeats/Cas.

Language: Английский

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Nanoparticle-Mediated Delivery towards Advancing Plant Genetic Engineering

Trends in biotechnology, Journal Year: 2018, Volume and Issue: 36(9), P. 882 - 897

Published: April 24, 2018

Language: Английский

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Non-viral delivery systems for CRISPR/Cas9-based genome editing: Challenges and opportunities

Biomaterials, Journal Year: 2018, Volume and Issue: 171, P. 207 - 218

Published: April 18, 2018

In recent years, CRISPR (clustered regularly interspaced short palindromic repeat)/Cas (CRISPR-associated) genome editing systems have become one of the most robust platforms in basic biomedical research and therapeutic applications. To date, efficient vivo delivery CRISPR/Cas9 system to targeted cells remains a challenge. Although viral vectors been widely used vitro vivo, their fundamental shortcomings, such as risk carcinogenesis, limited insertion size, immune responses difficulty large-scale production, severely limit further Alternative non-viral for are urgently needed. With rapid development vectors, lipid- or polymer-based nanocarriers shown great potential delivery. this review, we analyze pros cons delivering form plasmid, mRNA, protein then discuss limitations challenges CRISPR/Cas9-based editing. Furthermore, current that applied outlined details. Finally, critical obstacles highlighted promising strategies overcome these barriers proposed.

Language: Английский

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CRISPR-Cas12a-Based Nucleic Acid Amplification-Free DNA Biosensor via Au Nanoparticle-Assisted Metal-Enhanced Fluorescence and Colorimetric Analysis

Nano Letters, Journal Year: 2020, Volume and Issue: 21(1), P. 693 - 699

Published: Dec. 21, 2020

Cell-free DNA (cfDNA) has attracted significant attention due to its high potential diagnose diseases, such as cancer. Still, detection by amplification method limitations because of false-positive signals and difficulty in designing target-specific primers. CRISPR-Cas-based fluorescent biosensors have been developed but also need the step for detection. In this study, first time CRISPR-Cas12a based nucleic acid amplification-free biosensor was detect cfDNA a metal-enhanced fluorescence (MEF) using DNA-functionalized Au nanoparticle (AuNP). Upon activating complex target subsequent single-strand (ssDNA) degradation between AuNP fluorophore, MEF occurred with color changes from purple red-purple. Using system, breast cancer gene-1 (BRCA-1) can be detected very sensitivity 30 min. This rapid highly selective sensor applied measure other biomarkers viral field-deployable point-of-care testing (POCT) platform.

Language: Английский

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Strategies in the delivery of Cas9 ribonucleoprotein for CRISPR/Cas9 genome editing

Theranostics, Journal Year: 2020, Volume and Issue: 11(2), P. 614 - 648

Published: Oct. 28, 2020

CRISPR/Cas9 genome editing has gained rapidly increasing attentions in recent years, however, the translation of this biotechnology into therapy been hindered by efficient delivery materials target cells. Direct system as a ribonucleoprotein (RNP) complex consisting Cas9 protein and single guide RNA (sgRNA) emerged powerful widespread method for due to its advantages transient reduced off-target effects. In review, we summarized current RNP systems including physical approaches synthetic carriers. The mechanisms beneficial roles these strategies intracellular were reviewed. Examples development stimuli-responsive targeted carriers are highlighted. Finally, challenges perspectives rational design next generation promising field will be discussed.

Language: Английский

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CRISPR/Cas-Mediated Base Editing: Technical Considerations and Practical Applications

Trends in biotechnology, Journal Year: 2019, Volume and Issue: 37(10), P. 1121 - 1142

Published: April 14, 2019

Language: Английский

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Lipid-Based DNA Therapeutics: Hallmarks of Non-Viral Gene Delivery

ACS Nano, Journal Year: 2019, Volume and Issue: 13(4), P. 3754 - 3782

Published: March 25, 2019

Gene therapy is a promising strategy for the treatment of monogenic disorders. Non-viral gene delivery systems including lipid-based DNA therapeutics offer opportunity to deliver an encoding sequence specifically target tissue and thus enable expression therapeutic proteins in diseased cells. Currently, available approaches based on are inefficient require improvements achieve clinical utility. In this Review, we discuss state-of-the-art that have been investigated preclinical setting. We emphasize factors influencing subsequent vitro, ex vivo, vivo. addition, cover aspects nanoparticle engineering optimization therapeutics. Finally, highlight achievements therapies trials.

Language: Английский

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Treating Disease at the RNA Level with Oligonucleotides

New England Journal of Medicine, Journal Year: 2019, Volume and Issue: 380(1), P. 57 - 70

Published: Jan. 2, 2019

Interview with Dr. Arthur A. Levin on the use of oligonucleotide drugs that regulate RNA. (10:24)Download As research structure and function RNAs has rapidly expanded, so too number in clinical development This article describes several examples suggests where field is headed.

Language: Английский

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