Displaying publications 1 - 20 of 43 in total

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  1. A potential paradigm in CRISPR/Cas systems delivery: at the crossroad of microalgal gene editing and algal-mediated nanoparticles
    Feng S, Xie X, Liu J, Li A, Wang Q, Guo D, et al.
    J Nanobiotechnology, 2023 Oct 10;21(1):370.
    PMID: 37817254 DOI: 10.1186/s12951-023-02139-z
    Microalgae as the photosynthetic organisms offer enormous promise in a variety of industries, such as the generation of high-value byproducts, biofuels, pharmaceuticals, environmental remediation, and others. With the rapid advancement of gene editing technology, CRISPR/Cas system has evolved into an effective tool that revolutionised the genetic engineering of microalgae due to its robustness, high target specificity, and programmability. However, due to the lack of robust delivery system, the efficacy of gene editing is significantly impaired, limiting its application in microalgae. Nanomaterials have become a potential delivery platform for CRISPR/Cas systems due to their advantages of precise targeting, high stability, safety, and improved immune system. Notably, algal-mediated nanoparticles (AMNPs), especially the microalgae-derived nanoparticles, are appealing as a sustainable delivery platform because of their biocompatibility and low toxicity in a homologous relationship. In addition, living microalgae demonstrated effective and regulated distribution into specified areas as the biohybrid microrobots. This review extensively summarised the uses of CRISPR/Cas systems in microalgae and the recent developments of nanoparticle-based CRISPR/Cas delivery systems. A systematic description of the properties and uses of AMNPs, microalgae-derived nanoparticles, and microalgae microrobots has also been discussed. Finally, this review highlights the challenges and future research directions for the development of gene-edited microalgae.
    Matched MeSH terms: CRISPR-Cas Systems/genetics
  2. dCas9 Tells Tales: Probing Gene Function and Transcription Regulation in Cancer
    Mohamad Zamberi NN, Abuhamad AY, Low TY, Mohtar MA, Syafruddin SE
    CRISPR J, 2024 Apr;7(2):73-87.
    PMID: 38635328 DOI: 10.1089/crispr.2023.0078
    Clustered regularly interspaced short palindromic repeats (CRISPR)-based genome editing is evolving into an essential tool in the field of biological and medical research. Notably, the development of catalytically deactivated Cas9 (dCas9) enzyme has substantially broadened its traditional boundaries in gene editing or perturbation. The conjugation of dCas9 with various molecular effectors allows precise control over transcriptional processes, epigenetic modifications, visualization of chromosomal dynamics, and several other applications. This expanded repertoire of CRISPR-Cas9 applications has emerged as an invaluable molecular tool kit that empowers researchers to comprehensively interrogate and gain insights into health and diseases. This review delves into the advancements in Cas9 protein engineering, specifically on the generation of various dCas9 tools that have significantly enhanced the CRISPR-based technology capability and versatility. We subsequently discuss the multifaceted applications of dCas9, especially in interrogating the regulation and function of genes that involve in supporting cancer pathogenesis. In addition, we also delineate the designing and utilization of dCas9-based tools as well as highlighting its current constraints and transformative potentials in cancer research.
    Matched MeSH terms: CRISPR-Cas Systems/genetics
  3. Aptamer-CRISPR/Cas as a tool in point-of-care testing of infectious diseases
    Lau CH, Huang Y, Ch'ng ES, Tang TH, Zhu H, Citartan M
    Talanta, 2027 Mar 01;313(Pt A):130767.
    PMID: 42892010 DOI: 10.1016/j.talanta.2026.130767
    Aptamers and the CRISPR/Cas system have become the cornerstones of target detections due to their numerous advantages. Aptamers enable direct recognition of non-nucleic acid targets, such as pathogens and antigens, while CRISPR/Cas system drives signal amplification and rapid detection. The combined aptamer-CRISPR/Cas system can overcome the limitations of traditional target detection by realizing field-deployable, highly sensitive, specific, versatile, and rapid assays. Given the increasing number of different aptamer-CRISPR/Cas systems developed recently, particularly for infectious diseases, we provide a comprehensive overview of these platforms designed for detecting significant viruses such as SARS-CoV-2, Monkeypox virus, Hepatitis B virus and bacteria that include Mycobacterium tuberculosis, Staphylococcus aureus and Salmonella. Current challenges that impede the deployment of aptamer-CRISPR/Cas for point-of-care pathogen detection were also mapped, including primary barriers such as clinical matrix effect and stability. Several strategies to mitigate these issues were also set forth, predominantly the integration of artificial intelligence and machine learning to accelerate the transition of aptamer-CRISPR/Cas system to the clinical translation and commercialization phase, paving the way towards widespread implementation in global health.
    Matched MeSH terms: CRISPR-Cas Systems*
  4. Fulltext CRISPR/dCas9 platforms in plants: strategies and applications beyond genome editing
    Moradpour M, Abdulah SNA
    Plant Biotechnol J, 2020 Jan;18(1):32-44.
    PMID: 31392820 DOI: 10.1111/pbi.13232
    Clustered regularly interspaced short palindromic repeat (CRISPR) and Cas9-associated protein systems provide a powerful genetic manipulation tool that can drive plant research forward. Nuclease-dead Cas9 (dCas9) is an enzymatically inactive mutant of Cas9 in which its endonuclease activity is non-functional. The applications of CRISPR/dCas9 have expanded and diversified in recent years. Originally, dCas9 was used as a CRISPR/Cas9 re-engineering tool that enables targeted expression of any gene or multiple genes through recruitment of transcriptional effector domains without introducing irreversible DNA-damaging mutations. Subsequent applications have made use of its ability to recruit modifying enzymes and reporter proteins to DNA target sites. In this paper, the most recent progress in the applications of CRISPR/dCas9 in plants, which include gene activation and repression, epigenome editing, modulation of chromatin topology, live-cell chromatin imaging and DNA-free genetic modification, will be reviewed. The associated strategies for exploiting the CRISPR/dCas9 system for crop improvement with a dimer of the future of the CRISPR/dCas9 system in the functional genomics of crops and the development of traits will be briefly discussed.
    Matched MeSH terms: CRISPR-Cas Systems*
  5. CRISPR-Cas9 Genome Editing Tool for the Production of Industrial Biopharmaceuticals
    Khan AH, Tye GJ, Noordin R
    Mol Biotechnol, 2020 Sep;62(9):401-411.
    PMID: 32749657 DOI: 10.1007/s12033-020-00265-9
    A broad range of cell lines with characteristic features are used as bio-factories to produce recombinant proteins for basic research and therapeutic purposes. Genetic engineering strategies have been used to manipulate the genome of mammalian cells, insects, and yeasts for heterologous expression. One reason is that the glycosylation pattern of the expression hosts differs somehow from mammalian cells, which may cause immunogenic reactions upon administration in humans. CRISPR-Cas9 is a simple, efficient, and versatile genome engineering tool that can be programmed to precisely make double-stranded breaks at the desired loci. Compared to the classical genome editing methods, a CRISPR-Cas9 system is an ideal tool, providing the opportunity to integrate or delete genes from the target organisms. Besides broadened applications, limited studies have used CRISPR-Cas9 for editing the endogenous pathways in expression systems for biopharmaceutical applications. In the present review, we discuss the use of CRISPR-Cas9 in expression systems to improve host cell lines, increase product yield, and humanize glycosylation pathways by targeting intrinsic genes.
    Matched MeSH terms: CRISPR-Cas Systems*
  6. A review of CRISPR-Cas and PCR-based methods for the detection of animal species in the food chain-current challenges and future prospects
    Sultana S, Azlan A, Mohd Desa MN, Mahyudin NA, Anburaj A
    Food Addit Contam Part A Chem Anal Control Expo Risk Assess, 2024 Mar;41(3):213-227.
    PMID: 38284970 DOI: 10.1080/19440049.2024.2304577
    Regular testing and systematic investigation play a vital role to ensure product safety. Until now, the existing food authentication techniques have been based on proteins, lipids, and nucleic acid-based assays. Among various deoxyribonucleic acid (DNA)-based methods, the recently developed Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) based bio-sensing is an innovative and fast-expanding technology. The CRISPR/Cas-9 is known as Clustered Regularly Interspaced Short Palindromic Repeats due to the flexibility and simplicity of the CRISPR/Cas9 site-specific editing tool has been applied in many biological research areas such as Gene therapy, cell line development, discovering mechanisms of disease, and drug discovery. Nowadays, the CRISPR-Cas system has also been introduced into food authentication via detecting DNA barcodes of poultry and livestock both in processed and unprocessed food samples. This review documents various DNA based approaches, in an accessible format. Future CRISPR technologies are forecast while challenges are outlined.
    Matched MeSH terms: CRISPR-Cas Systems*
  7. Recent trends in cancer therapy: A review on the current state of gene delivery
    Yahya EB, Alqadhi AM
    Life Sci, 2021 Mar 15;269:119087.
    PMID: 33476633 DOI: 10.1016/j.lfs.2021.119087
    Cancer treatment has been always considered one of the most critical and vital themes of clinical issues. Many approaches have been developed, depending on the type and the stage of tumor. Gene therapy has the potential to revolutionize different cancer therapy. With the advent of recent bioinformatics technologies and genetic science, it become possible to identify, diagnose and determine the potential treatment using the technology of gene delivery. Several approaches have been developed and experimented in vitro and vivo for cancer therapy including: naked nucleic acids based therapy, targeting micro RNAs, oncolytic virotherapy, suicide gene based therapy, targeting telomerase, cell mediated gene therapy, and CRISPR/Cas9 based therapy. In this review, we present a straightforward introduction to cancer biology and occurrence, highlighting different viral and non-viral gene delivery systems for gene therapy and critically discussed the current and various strategies for cancer gene therapy.
    Matched MeSH terms: CRISPR-Cas Systems*
  8. Dual-Functional Tetrahedron Multivalent Aptamer Assisted Amplification-Free CRISPR/Cas12a Assay for Sensitive Detection of Salmonella
    Qiao Z, Xue L, Sun M, Ma N, Shi H, Yang W, et al.
    J Agric Food Chem, 2024 Jan 10;72(1):857-864.
    PMID: 38134022 DOI: 10.1021/acs.jafc.3c07582
    Salmonellosis continues to impose a significant economic burden globally. Rapid and sensitive detection of Salmonella is crucial to preventing the outbreaks of foodborne illnesses, yet it remains a formidable challenge. Herein, a dual-functional tetrahedron multivalent aptamer assisted amplification-free CRISPR/Cas12a assay was developed for Salmonella detection. In the system, the aptamer was programmatically assembled on the tetrahedral DNA nanostructure to fabricate a multivalent aptamer (TDN-multiApt), which displayed a 3.5-fold enhanced avidity over the monovalent aptamer and possessed four CRISPR/Cas12a targeting fragments to amplify signal. Therefore, TDN-multiApt could directly activate Cas12a to achieve the second signal amplification without any nucleic acid amplification. By virtue of the synergism of high avidity and cascaded signal amplifications, the proposed method allowed the ultrasensitive detection of Salmonella as low as 7 cfu mL-1. Meanwhile, this novel platform also exhibited excellent specificity against target bacteria and performed well in the detection of various samples, indicating its potential application in real samples.
    Matched MeSH terms: CRISPR-Cas Systems*
  9. The Promises and Pitfalls of CRISPR-Mediated Base Editing in Stem Cells
    Wong PK, Mohamad Zamberi NN, Syafruddin SE, Cheah FC, Azmi N, Law JX, et al.
    CRISPR J, 2023 Jun;6(3):196-215.
    PMID: 37219623 DOI: 10.1089/crispr.2023.0013
    Stem cells such as induced pluripotent stem cells, embryonic stem cells, and hematopoietic stem and progenitor cells are growing in importance in disease modeling and regenerative medicine. The applications of CRISPR-based gene editing to create a mélange of disease and nondisease stem cell lines have further enhanced the utility of this innately versatile group of cells in the studies of human genetic disorders. Precise base edits can be achieved using a variety of CRISPR-centric approaches, particularly homology-directed repair and the recently developed base editors and prime editors. Despite its much-touted potential, editing single DNA bases is technically challenging. In this review, we discuss the strategies for achieving exact base edits in the creation of various stem cell-based models for use in elucidating disease mechanisms and assessing drug efficacy, and the unique characteristics of stem cells that warrant special considerations.
    Matched MeSH terms: CRISPR-Cas Systems/genetics
  10. Studying the cellular efficacy and tolerability of using CRISPR-gRNA ribonucleoprotein (RNP) complex for in-vitro knockdown of TRIB2 in acute myeloid leukaemia (AML) cells as preliminary clinical evaluations
    Tan JYP, Thevendran R, Quek YS, Maheswaran S
    Mol Biol Rep, 2025 Apr 09;52(1):379.
    PMID: 40205014 DOI: 10.1007/s11033-025-10487-2
    BACKGROUND: Acute Myeloid Leukemia (AML) is a complex, genetically driven cancer disease with several mutations that complicate therapy regimes. The Tribbles gene family, specifically the TRIB2 gene, has garnered substantial interest as a crucial oncogenic factor of AML progression. Studying cancer through gene expression studies and focusing on the primary oncogenes provide accurate information for future therapies. Here, CRISPR stands as the most exceptional tool used to explore both oncogene functionalities and therapeutic utilities. The ribonucleoprotein (RNP) mode of CRISPR formation and usage are preferred compared to plasmid-mediated CRISPR systems, however, it can lead to complications post-transfection to sensitive cellular entities such as human cancer cells in-vitro, transcribing to similar outcomes in-vivo as well. Therefore, this study describes the use of in-house designed CRISPR-RNP systems targeting the TRIB2 oncogene and evaluates their post-transfection cellular safety and efficacy aspects for future clinical applications.

    METHODS: This study uses a designed guide RNA targeting the TRIB2 gene assembled via In-vitro synthesis. The gRNA with Cas9 protein leads to the formation of CRISPR-RNP structures, which target and cleave the TRIB2 gene. The assembled CRISPR-RNP system is transfected into target AML cell and control cell lines (i.e. HEK cells), and the subsequent gene cleavage and resulting changes to the AML cells in terms of cellular safety/tolerability and gene knockdown efficacy were studied via RT-qPCR, flow cytometry, and cell viability analysis.

    RESULTS: The outcome demonstrates the well-tolerated transfection of the in-vitro assembled CRISPR RNP system with no signs of cellular toxicity and disruptions towards the AML cell's metabolic activities, promoting the safety aspects of CRISPR RNP post-transfection to human cells. The study further highlights the in-vitro efficacy of the CRISPR RNP in targeting the TRIB2 oncogene, where a statistically significant gene knockdown of more than 80% was detected via qPCR analysis of TRIB2 gene expression with minimal to no background effects from individual RNP components, equating to their targeted gene cleavage effects. In addition, the CRISPR TRIB2 gene knockdown also indicated the possibilities of induced AML cell death measured via flow cytometry markers, translating to favourable outcomes in eliminating cancerous growths.

    CONCLUSIONS: This study research contributes to the refinement of the CRISPR-RNP strategy and preliminary evaluation for future clinical uses.

    Matched MeSH terms: CRISPR-Cas Systems/genetics
  11. CRISPRi-mediated silencing of efflux pump genes restores antibiotic susceptibility in multidrug-resistant Acinetobacter baumannii
    Zainul R, Abdul Khaleq MA, Farah H, Panigrahi R, Muniyandy E, Kumar N, et al.
    Microb Pathog, 2026 Dec;221:108854.
    PMID: 42810556 DOI: 10.1016/j.micpath.2026.108854
    Multidrug efflux pumps are a critical mechanism underlying both intrinsic and acquired resistance in Acinetobacter baumannii. One of the most perilous multidrug-resistant (MDR) bacterial infections. Resistance-nodulation-division transporters, including AdeABC, AdeIJK, and AdeFGH, are critical efflux systems that diminish intracellular antibiotic levels, enhance bacterial persistence, support biofilm-related tolerance, and lead to treatment failure. Recent advancements in CRISPR interference (CRISPRi), a programmable gene-silencing method using catalytically inactive Cas9 (dCas9), have enabled a versatile platform for reversible, sequence-specific transcriptional repression without inducing double-strand DNA breaks. CRISPRi has emerged as an essential instrument for functional genomics, antimicrobial target validation, and the investigation of complex resistance networks in multidrug-resistant bacteria because to its precision, adaptability, and multiplexing capacities. This work primarily focuses on the recent advancements in CRISPRi-mediated silencing of efflux pump genes and their regulatory circuits in A. baumannii. Targeted suppression of key RND transporters, including adeB, adeJ, and adeG, along with regulatory components such as adeRS, has demonstrated in experiments a substantial decrease in efflux activity, an increase in intracellular antibiotic accumulation, a reduction in minimum inhibitory concentrations, and a reinstatement of susceptibility to various antimicrobial classes, including carbapenems, fluoroquinolones, aminoglycosides, tetracyclines, tigecycline, and chloramphenicol. CRISPRi-based functional screening has identified weaknesses in lipooligosaccharide synthesis, outer membrane biogenesis, and global regulatory pathways, which together augment antibiotic effectiveness with direct efflux pump inhibition. Recent research suggests that the multifactorial resistance mechanisms of MDR A. baumannii may be addressed by multiplex CRISPRi methods and combination therapies that include gene silencing with conventional antibiotics or efflux pump inhibitors. The evaluation also critically examines current delivery techniques, which will eventually determine the feasibility of translating CRISPRi medicines. Examples of contemporary platforms include plasmid-based systems, chromosomally integrated expression cassettes, conjugation-mediated transfer, bacteriophage-mediated delivery, and innovative nanotechnology-assisted carriers designed to improve intracellular delivery and bacterial targeting. Despite substantial advancements in experimental CRISPRi methodologies, the efficient delivery to infected tissues and clinical isolates remains a considerable challenge. Challenges include restricted infiltration into established biofilms, variable gene repression among genetically diverse strains, potential off-target effects, emergence of escape mutations, preservation of genetic stability, immune-mediated elimination of delivery vehicles, large-scale production and regulatory hurdles, and the absence of dependable animal and clinical studies validating therapeutic efficacy and safety. Current knowledge indicates that CRISPRi-mediated efflux pump silencing is a very promising precision antimicrobial adjuvant method that might enhance the efficacy of existing antibiotics against MDR A. baumannii. Before this technique can be integrated into regular clinical practice, substantial technological advancements, improved delivery systems, comprehensive biosafety evaluations, and rigorous in vivo and clinical validations will be required.
    Matched MeSH terms: CRISPR-Cas Systems*
  12. Fulltext Efficiency of genome editing using modified single-stranded oligodeoxyribonucleotides in human cells
    Kim S, Matsushita Y, Katagiri T, Maseda H
    Sci Rep, 2025 Mar 21;15(1):9764.
    PMID: 40119107 DOI: 10.1038/s41598-025-94071-5
    Single-stranded oligodeoxyribonucleotide (ssODN) gene editing has emerged as a promising therapeutic strategy. However, further improvements in efficiency are desired for practical application. The effects of strand length and locked nucleic acid (LNA) modification on ssODN genome editing were investigated by introducing an assay cassette into the genome of HEK293T cells and measuring precise base deletions of eight bases. The introduction of LNAs into ssODNs, five pairs of LNAs at 25-35 nt from the centre and one pair at 20-25 nt, showed approximately 18-fold higher efficiency than unmodified ssODNs of the same length in the study using 70 nt ssODNs. In addition, genome editing efficiency was further improved when LNAs were introduced at the same positions as the 70 nt ssODN, which showed the highest efficiency for the 90 nt ssODN. However, in some cases, the same number of LNA modifications could conversely reduce the efficiency, and the modification positions in the ssODN method were successfully optimised in the present study. Furthermore, the oligo DNA was shown to be effective not only for deletions but also for base substitutions, with an editing efficiency of 0.63% per cell.
    Matched MeSH terms: CRISPR-Cas Systems
  13. Knockout of CTNNB1 by CRISPR-Cas9 technology inhibits cell proliferation through the Wnt/β-catenin signaling pathway
    Guan L, Zhu S, Han Y, Yang C, Liu Y, Qiao L, et al.
    Biotechnol Lett, 2018 Mar;40(3):501-508.
    PMID: 29249062 DOI: 10.1007/s10529-017-2491-2
    OBJECTIVE: To study the effects of CTNNB1 gene knockout by CRISPR-Cas9 technology on cell adhesion, proliferation, apoptosis, and Wnt/β-catenin signaling pathway.

    RESULTS: CTNNB1 gene of HEK 293T cells was knocked out by CRISPR-Cas9. This was confirmed by sequencing and western blotting. Methylthiazolyl-tetrazolium bromide assays indicated that deletion of β-catenin significantly weakened adhesion ability and inhibited proliferation rate (P 

    Matched MeSH terms: CRISPR-Cas Systems/genetics*
  14. Fulltext The phylogenomics of CRISPR-Cas system and revelation of its features in Salmonella
    Kushwaha SK, Bhavesh NLS, Abdella B, Lahiri C, Marathe SA
    Sci Rep, 2020 12 03;10(1):21156.
    PMID: 33273523 DOI: 10.1038/s41598-020-77890-6
    Salmonellae display intricate evolutionary patterns comprising over 2500 serovars having diverse pathogenic profiles. The acquisition and/or exchange of various virulence factors influences the evolutionary framework. To gain insights into evolution of Salmonella in association with the CRISPR-Cas genes we performed phylogenetic surveillance across strains of 22 Salmonella serovars. The strains differed in their CRISPR1-leader and cas operon features assorting into two main clades, CRISPR1-STY/cas-STY and CRISPR1-STM/cas-STM, comprising majorly typhoidal and non-typhoidal Salmonella serovars respectively. Serovars of these two clades displayed better relatedness, concerning CRISPR1-leader and cas operon, across genera than between themselves. This signifies the acquisition of CRISPR1/Cas region could be through a horizontal gene transfer event owing to the presence of mobile genetic elements flanking CRISPR1 array. Comparison of CRISPR and cas phenograms with that of multilocus sequence typing (MLST) suggests differential evolution of CRISPR/Cas system. As opposed to broad-host-range, the host-specific serovars harbor fewer spacers. Mapping of protospacer sources suggested a partial correlation of spacer content with habitat diversity of the serovars. Some serovars like serovar Enteritidis and Typhimurium that inhabit similar environment/infect similar hosts hardly shared their protospacer sources.
    Matched MeSH terms: CRISPR-Cas Systems/genetics*
  15. Fulltext Harnessing the Potential of CRISPR/Cas in Atherosclerosis: Disease Modeling and Therapeutic Applications
    Siew WS, Tang YQ, Kong CK, Goh BH, Zacchigna S, Dua K, et al.
    Int J Mol Sci, 2021 Aug 05;22(16).
    PMID: 34445123 DOI: 10.3390/ijms22168422
    Atherosclerosis represents one of the major causes of death globally. The high mortality rates and limitations of current therapeutic modalities have urged researchers to explore potential alternative therapies. The clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9) system is commonly deployed for investigating the genetic aspects of Atherosclerosis. Besides, advances in CRISPR/Cas system has led to extensive options for researchers to study the pathogenesis of this disease. The recent discovery of Cas9 variants, such as dCas9, Cas9n, and xCas9 have been established for various applications, including single base editing, regulation of gene expression, live-cell imaging, epigenetic modification, and genome landscaping. Meanwhile, other Cas proteins, such as Cas12 and Cas13, are gaining popularity for their applications in nucleic acid detection and single-base DNA/RNA modifications. To date, many studies have utilized the CRISPR/Cas9 system to generate disease models of atherosclerosis and identify potential molecular targets that are associated with atherosclerosis. These studies provided proof-of-concept evidence which have established the feasibility of implementing the CRISPR/Cas system in correcting disease-causing alleles. The CRISPR/Cas system holds great potential to be developed as a targeted treatment for patients who are suffering from atherosclerosis. This review highlights the advances in CRISPR/Cas systems and their applications in establishing pathogenetic and therapeutic role of specific genes in atherosclerosis.
    Matched MeSH terms: CRISPR-Cas Systems/genetics*
  16. The Smart Programmable CRISPR Technology: A Next Generation Genome Editing Tool for Investigators
    Chakraborty C, Teoh SL, Das S
    Curr Drug Targets, 2017;18(14):1653-1663.
    PMID: 27231109 DOI: 10.2174/1389450117666160527142321
    BACKGROUND: The present era is fast experiencing rapid innovation in the genome-editing technology. CRISPR Cas9-mediated targeted genetic manipulation is an easy, cost-effective and scalable method. As a result, it can be used for a broad range of targeted genome engineering.

    OBJECTIVE: The main objective of the present review is to highlight the structural signature, classification, its mechanism and application from basic science to medicine and future challenges for this genome editing tool kit.

    RESULTS: The present review provides a brief description of the recent development of CRISPR-Cas9 genome editing technology. We discuss the paradigms shift for this next generation genome editing technology, CRISPR. The CRISPR structural significance, classification and its different applications are also being discussed. We portray the future challenges for this extraordinary genome in vivo editing tool. We also highlight the role of CRISPR genome editing in curing many diseases.

    CONCLUSION: Scientists and researchers are constantly looking one genome editing tool that is competent, simple and low-cost assembly of nucleases. It can target any particular site without any off-target mutations in the genome. The CRISPR-Cas9 has all of the above characteristics. The genome engineering technology may be a strong and inspiring technology meant for the next generation of drug development.

    Matched MeSH terms: CRISPR-Cas Systems*
  17. Fulltext Therapeutic potentials of CRISPR-Cas genome editing technology in human viral infections
    Najafi S, Tan SC, Aghamiri S, Raee P, Ebrahimi Z, Jahromi ZK, et al.
    Biomed Pharmacother, 2022 Apr;148:112743.
    PMID: 35228065 DOI: 10.1016/j.biopha.2022.112743
    Viral infections are a common cause of morbidity worldwide. The emergence of Coronavirus Disease 2019 (COVID-19) has led to more attention to viral infections and finding novel therapeutics. The CRISPR-Cas9 system has been recently proposed as a potential therapeutic tool for the treatment of viral diseases. Here, we review the research progress in the use of CRISPR-Cas technology for treating viral infections, as well as the strategies for improving the delivery of this gene-editing tool in vivo. Key challenges that hinder the widespread clinical application of CRISPR-Cas9 technology are also discussed, and several possible directions for future research are proposed.
    Matched MeSH terms: CRISPR-Cas Systems*
  18. Fulltext A redefined InDel taxonomy provides insights into mutational signatures
    Koh GCC, Nanda AS, Rinaldi G, Boushaki S, Degasperi A, Badja C, et al.
    Nat Genet, 2025 May;57(5):1132-1141.
    PMID: 40210680 DOI: 10.1038/s41588-025-02152-y
    Despite their deleterious effects, small insertions and deletions (InDels) have received far less attention than substitutions. Here we generated isogenic CRISPR-edited human cellular models of postreplicative repair dysfunction (PRRd), including individual and combined gene edits of DNA mismatch repair (MMR) and replicative polymerases (Pol ε and Pol δ). Unique, diverse InDel mutational footprints were revealed. However, the prevailing InDel classification framework was unable to discriminate these InDel signatures from background mutagenesis and from each other. To address this, we developed an alternative InDel classification system that considers flanking sequences and informative motifs (for example, longer homopolymers), enabling unambiguous InDel classification into 89 subtypes. Through focused characterization of seven tumor types from the 100,000 Genomes Project, we uncovered 37 InDel signatures; 27 were new. In addition to unveiling previously hidden biological insights, we also developed PRRDetect-a highly specific classifier of PRRd status in tumors, with potential implications for immunotherapies.
    Matched MeSH terms: CRISPR-Cas Systems/genetics
  19. Fulltext CRISPR/Cas9 mediated ENT2 gene knockout altered purine catabolic pathway and induced apoptosis in colorectal cell lines
    Naes SM, Ab-Rahim S, Mazlan M, Syafruddin SE, Mohtar MA, Abuhamad AY, et al.
    PLoS One, 2025;20(8):e0329501.
    PMID: 40824957 DOI: 10.1371/journal.pone.0329501
    Although purine metabolism is one of the most impacted pathways in colorectal cancer (CRC), little is known about the role of equilibrative nucleoside transporter 2 (ENT2) in CRC development and its association with the altered purine metabolism pathway. This study aimed to determine the role of ENT2 in altered purine metabolism in the early and late stages of CRC using CRISPR/Cas9 gene editing tools and a variety of functional experiments. The expression of ENT2 was significantly higher (P CRISPR/Cas9 tool. The hypoxanthine (HPX) level and the xanthine oxidase (XO) activity were significantly higher in both HT29/KO and DLD1/KO single cell-derived clones (P 
    Matched MeSH terms: CRISPR-Cas Systems*
  20. Fulltext The Impact of CRISPR-Cas System on Antiviral Therapy
    Bayat H, Naderi F, Khan AH, Memarnejadian A, Rahimpour A
    Adv Pharm Bull, 2018 Nov;8(4):591-597.
    PMID: 30607331 DOI: 10.15171/apb.2018.067
    Clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein nuclease (Cas) is identified as an adaptive immune system in archaea and bacteria. Type II of this system, CRISPR-Cas9, is the most versatile form that has enabled facile and efficient targeted genome editing. Viral infections have serious impacts on global health and conventional antiviral therapies have not yielded a successful solution hitherto. The CRISPR-Cas9 system represents a promising tool for eliminating viral infections. In this review, we highlight 1) the recent progress of CRISPR-Cas technology in decoding and diagnosis of viral outbreaks, 2) its applications to eliminate viral infections in both pre-integration and provirus stages, and 3) various delivery systems that are employed to introduce the platform into target cells.
    Matched MeSH terms: CRISPR-Cas Systems
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