Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing

Biologics, Journal Year: 2021, Volume and Issue: Volume 15, P. 353 - 361

Published: Aug. 1, 2021

Abstract: Clustered regularly interspaced short palindromic repeat (CRISPR) and their associated protein (Cas-9) is the most effective, efficient, accurate method of genome editing tool in all living cells utilized many applied disciplines. Guide RNA (gRNA) CRISPR-associated proteins are two essential components CRISPR/Cas-9 system. The mechanism contains three steps, recognition, cleavage, repair. designed sgRNA recognizes target sequence gene interest through a complementary base pair. While Cas-9 nuclease makes double-stranded breaks at site 3 pair upstream to protospacer adjacent motif, then break repaired by either non-homologous end joining or homology-directed repair cellular mechanisms. genome-editing has wide number applications areas including medicine, agriculture, biotechnology. In it could help design new grains improve nutritional value. being investigated for cancers, HIV, therapy such as sickle cell disease, cystic fibrosis, Duchenne muscular dystrophy. technology also regulation specific genes advanced modification protein. However, immunogenicity, effective delivery systems, off-target effect, ethical issues have been major barriers extend clinical applications. Although becomes era molecular biology countless roles ranging from basic researches applications, there still challenges rub practical various improvements needed overcome obstacles. Keywords: CRISPR, Cas-9, sgRNA, gene-editing, mechanism,

Language: Английский

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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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In vivo delivery of CRISPR-Cas9 using lipid nanoparticles enables antithrombin gene editing for sustainable hemophilia A and B therapy

Science Advances, Journal Year: 2022, Volume and Issue: 8(3)

Published: Jan. 21, 2022

Hemophilia is a hereditary disease that remains incurable. Although innovative treatments such as gene therapy or bispecific antibody have been introduced, substantial unmet needs still exist with respect to achieving long-lasting therapeutic effects and treatment options for inhibitor patients. Antithrombin (AT), an endogenous negative regulator of thrombin generation, potent genome editing target sustainable patients hemophilia A B. In this study, we developed optimized lipid nanoparticles (LNPs) deliver Cas9 mRNA along single guide RNA targeted AT in the mouse liver. The LNP-mediated CRISPR-Cas9 delivery resulted inhibition led improvement generation. Bleeding-associated phenotypes were recovered both B mice. No active off-targets, liver-induced toxicity, anti-Cas9 immune responses detected, indicating was safe efficient approach therapy.

Language: Английский

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Advances in CRISPR therapeutics

Nature Reviews Nephrology, Journal Year: 2022, Volume and Issue: 19(1), P. 9 - 22

Published: Oct. 24, 2022

Language: Английский

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Stimuli-Responsive Gene Delivery Nanocarriers for Cancer Therapy

Nano-Micro Letters, Journal Year: 2023, Volume and Issue: 15(1)

Published: Feb. 8, 2023

Gene therapy provides a promising approach in treating cancers with high efficacy and selectivity few adverse effects. Currently, the development of functional vectors safety effectiveness is intense focus for improving delivery nucleic acid drugs gene therapy. For this purpose, stimuli-responsive nanocarriers displayed strong potential overall efficiencies reducing effects via effective protection, prolonged blood circulation, specific tumor accumulation, controlled release profile drugs. Besides, synergistic could be achieved when combined other therapeutic regimens. This review summarizes recent advances various delivery. Particularly, responding to endogenous stimuli including pH, reactive oxygen species, glutathione, enzyme, etc., exogenous light, thermo, ultrasound, magnetic field, are introduced. Finally, future challenges prospects toward clinical translation well discussed. The major objective present biomedical cancer provide guidance developing novel nanoplatforms that clinically applicable.

Language: Английский

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Massively parallel base editing to map variant effects in human hematopoiesis

Cell, Journal Year: 2023, Volume and Issue: 186(11), P. 2456 - 2474.e24

Published: May 1, 2023

Systematic evaluation of the impact genetic variants is critical for study and treatment human physiology disease. While specific mutations can be introduced by genome engineering, we still lack scalable approaches that are applicable to important setting primary cells, such as blood immune cells. Here, describe development massively parallel base-editing screens in hematopoietic stem progenitor Such enable functional variant effects across any differentiation state. Moreover, they allow rich phenotyping through single-cell RNA sequencing readouts separately characterization editing outcomes pooled genotyping. We efficiently design improved leukemia immunotherapy approaches, comprehensively identify non-coding modulating fetal hemoglobin expression, define mechanisms regulating differentiation, probe pathogenicity uncharacterized disease-associated variants. These strategies will advance effective high-throughput variant-to-function mapping hematopoiesis causes diverse diseases.

Language: Английский

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Current State of Human Gene Therapy: Approved Products and Vectors

Pharmaceuticals, Journal Year: 2023, Volume and Issue: 16(10), P. 1416 - 1416

Published: Oct. 5, 2023

In the realm of gene therapy, a pivotal moment arrived with Paul Berg’s groundbreaking identification first recombinant DNA in 1972. This achievement set stage for future breakthroughs. Conditions once considered undefeatable, like melanoma, pancreatic cancer, and host other ailments, are now being addressed at their root cause—the genetic level. Presently, therapy landscape stands adorned 22 approved vivo ex products, including IMLYGIC, LUXTURNA, Zolgensma, Spinraza, Patisiran, many more. this comprehensive exploration, we delve into rich assortment 16 drugs, from siRNA, miRNA, CRISPR/Cas9 to aptamers TRAIL/APO2L, as well 46 carriers, AAV, AdV, LNPs, exosomes naked mRNA, sonoporation, magnetofection. The article also discusses advantages disadvantages each product vector type, current challenges faced practical use its potential.

Language: Английский

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Current and future concepts for the generation and application of genetically engineered CAR-T and TCR-T cells

Frontiers in Immunology, Journal Year: 2023, Volume and Issue: 14

Published: March 6, 2023

Adoptive cell therapy (ACT) has seen a steep rise of new therapeutic approaches in its immune-oncology pipeline over the last years. This is great part due to recent approvals chimeric antigen receptor (CAR)-T therapies and their remarkable efficacy certain soluble tumors. A big focus ACT lies on T cells how genetically modify them target kill tumor cells. Genetically modified that are currently utilized either equipped with an engineered CAR or (TCR) for this purpose. Both strategies have advantages limitations. While CAR-T already used clinic, these face challenges when it comes treatment solid New designs next-generation might be able overcome hurdles. Moreover, CARs restricted surface antigens. TCR-T targeting intracellular antigens provide necessary qualities In review, we will summarize major advancements technology. cover ongoing clinical trials, discuss current challenges, assessment future directions within field.

Language: Английский

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Recent advances in the delivery and applications of nonviral CRISPR/Cas9 gene editing

Drug Delivery and Translational Research, Journal Year: 2023, Volume and Issue: 13(5), P. 1500 - 1519

Published: March 29, 2023

Language: Английский

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Engineering approaches for RNA-based and cell-based osteoarthritis therapies

Nature Reviews Rheumatology, Journal Year: 2024, Volume and Issue: 20(2), P. 81 - 100

Published: Jan. 22, 2024

Language: Английский

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