CRISPR/Cas9 Delivery System Engineering for Genome Editing in Therapeutic Applications

Cheng, Hao; Zhang, Feng; Ding, Yang

doi:10.3390/pharmaceutics13101649

Cited by 45 publications

(46 citation statements)

References 71 publications

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“…However, an optimal deliver strategy to introduce the CRISPR-Cas9 system into the target bacterial populations remains a great challenge for plasmid or resistance gene elimination in the clinical application. 37 Most of the previous studies used physical or chemical methods, such as electroporation and chemical transformation, but these approaches are restricted to the Laboratory research and are not be applied in the clinical application. Considering that bacterial conjugation is critical for the dissemination of antimicrobial resistance, we aimed to explore conjugative plasmids to eliminate antibiotic resistance plasmids in bacteria.…”

Section: Discussionmentioning

confidence: 99%

Targeted Elimination of blaNDM-5 Gene in Escherichia coli by Conjugative CRISPR-Cas9 System

Li¹,

Wan²,

Zhao³

et al. 2022

IDR

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Purpose Plasmid-borne carbapenem resistance gene bla NDM-5 accelerates the dissemination of carbapenem-resistant Enterobacteriaceae. To efficiently eliminate the bla NDM-5 -harboring plasmid and sensitize the antibiotic-resistant bacteria to meropenem, we used the CRISPR-Cas9 system for combating the carbapenem-resistant Escherichia coli ( E. coil ). Methods A series of CRISPR-Cas9 plasmids was constructed, and specific guide RNAs(sgRNA) were designed to target the bla NDM-5 gene. We used chemically transformation or conjugation delivery methods, and the elimination efficiency in each recipient strains was evaluated by plate counting, PCR and quantitative real-time PCR (qPCR). Antimicrobial susceptibility test was carried out by using the broth microdilution method. In addition, we assessed the effect of the CRISPR-Cas9 system of adaptive immunity on the prevention of the exogenous resistant plasmids pNDM-5 by introducing the system into E coli J53. Results The results showed that pCas9, pCas9-oriT and pBAD-Cas9-oriT can effectively eliminate bla NDM-5 in E. coli with >94.00% elimination efficiency. The bla NDM-5 -harboring E. coli successfully restored their susceptibility to meropenem, with eight-fold reduction of minimum inhibitory concentration (MIC) values (from 16 µg/mL to 0.06 µg/mL). The E. coli J53 strain containing plasmid pCas9-N reduced the number of transconjugants by 26-fold. Conclusion The CRISPR-Cas9 system achieved plasmid clearance and simultaneous re-sensitization to meropenem in E. coli . The CRISPR-Cas9 system could block the horizontal transfer of plasmid pNDM-5. The conjugative delivery of CRISPR-Cas9 provides a new tool for the removal of resistance plasmids and sensitize the recipient to carbapenem. It provides a therapeutic approach to counteract the propagation of bla NDM-5 gene among clinical pathogens.

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Section: Discussionmentioning

confidence: 99%

Targeted Elimination of blaNDM-5 Gene in Escherichia coli by Conjugative CRISPR-Cas9 System

Li¹,

Wan²,

Zhao³

et al. 2022

IDR

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“…Continuous efforts in developing this science have provided excellent in vivo, in vitro, and ex vivo gene editing using diverse delivery strategies [162]. Importantly, this also has emerged as a powerful tool to manipulate the genome for therapeutic purposes to treat various genetic disorders such as thalassemia, tyrosinemia, or cancers [163]. In order to successfully deliver the CRISPR-Cas9 genome editing system, both physical techniques and delivery vectors are usually used.…”

Section: Delivery Of Crispr-casmentioning

confidence: 99%

CRISPR-Cas9-Based Technology and Its Relevance to Gene Editing in Parkinson’s Disease

et al. 2022

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Parkinson’s disease (PD) and other chronic and debilitating neurodegenerative diseases (NDs) impose a substantial medical, emotional, and financial burden on individuals and society. The origin of PD is unknown due to a complex combination of hereditary and environmental risk factors. However, over the last several decades, a significant amount of available data from clinical and experimental studies has implicated neuroinflammation, oxidative stress, dysregulated protein degradation, and mitochondrial dysfunction as the primary causes of PD neurodegeneration. The new gene-editing techniques hold great promise for research and therapy of NDs, such as PD, for which there are currently no effective disease-modifying treatments. As a result, gene therapy may offer new treatment options, transforming our ability to treat this disease. We present a detailed overview of novel gene-editing delivery vehicles, which is essential for their successful implementation in both cutting-edge research and prospective therapeutics. Moreover, we review the most recent advancements in CRISPR-based applications and gene therapies for a better understanding of treating PD. We explore the benefits and drawbacks of using them for a range of gene-editing applications in the brain, emphasizing some fascinating possibilities.

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“…There are various formats for the CRISPR/Cas9 system, including 1) ribonucleoprotein complexes containing Cas9 endonuclease protein and sgRNA molecule, 2) a plasmid that encodes both Cas9 and sgRNA, 3) two separate plasmids each encoding Cas9 and sgRNA, and 4) Cas9 mRNA and sgRNA molecule ( Table 2 ) ( Deng et al, 2019 ; Cheng et al, 2021 ).…”

Section: Types and Classes Crispr/cas Systemsmentioning

confidence: 99%

Nanocarriers: A novel strategy for the delivery of CRISPR/Cas systems

et al. 2022

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In recent decades, clustered regularly interspaced short palindromic repeat/CRISPR-associated protein (CRISPR/Cas) has become one of the most promising genome-editing tools for therapeutic purposes in biomedical and medical applications. Although the CRISPR/Cas system has truly revolutionized the era of genome editing, the safe and effective delivery of CRISPR/Cas systems represents a substantial challenge that must be tackled to enable the next generation of genetic therapies. In addition, there are some challenges in the in vivo delivery to the targeted cells/tissues. Nanotechnology-based drug delivery systems can be employed to overcome this issue. This review discusses different types and forms of CRISPR/Cas systems and the current CRISPR/Cas delivery systems, including non-viral carriers such as liposomes, polymeric, and gold particles. The focus then turns to the viral nanocarriers which have been recently used as a nanocarrier for CRISPR/Cas delivery.

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CRISPR/Cas9 Delivery System Engineering for Genome Editing in Therapeutic Applications

Cited by 45 publications

References 71 publications

Targeted Elimination of blaNDM-5 Gene in Escherichia coli by Conjugative CRISPR-Cas9 System

Targeted Elimination of blaNDM-5 Gene in Escherichia coli by Conjugative CRISPR-Cas9 System

CRISPR-Cas9-Based Technology and Its Relevance to Gene Editing in Parkinson’s Disease

Nanocarriers: A novel strategy for the delivery of CRISPR/Cas systems

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