Clustered Regularly Interspaced Short Palindromic Repeats Technology Research Articles

Advancements in CRISPR-Based Therapies for Genetic Modulation in Neurodegenerative Disorders.

: Neurodegenerative disorders pose significant challenges in the realm of healthcare, as these conditions manifest in complex, multifaceted ways, often attributed to genetic anomalies. With the emergence of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) technology, a new frontier has been unveiled in the quest for targeted, precise genetic manipulation. This abstract explores the recent advancements and potential applications of CRISPR-based therapies in addressing genetic components contributing to various neurodegenerative disorders. The review delves into the foundational principles of CRISPR technology, highlighting its unparalleled ability to edit genetic sequences with unprecedented precision. In addition, it talks about the latest progress in using CRISPR to target specific genetic mutations linked to neurodegenerative diseases like Huntington's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), and Parkinson's disease. It talks about the most important studies and trials that show how well and safely CRISPR-based therapies work. This shows how this technology can change genetic variants that cause diseases. Notably, the discussion emphasizes the challenges and ethical considerations associated with the implementation of CRISPR in clinical settings, including off-target effects, delivery methods, and long-term implications. Furthermore, the article explores the prospects and potential hurdles in the widespread application of CRISPR technology for treating neurodegenerative disorders. It touches upon the need for continued research, improved delivery mechanisms, and ethical frameworks to ensure responsible and equitable access to these groundbreaking therapies.

Assessing the environmental contribution of clustered regularly interspaced short palindromic repeats (CRISPR) rice in the presence of insect pest uncertainty

AbstractAdopting genome editing with the trait of pest resistance contributes to sustainable development by reducing pesticide use. Developed by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology, CRISPR rice is resistant to two of its most destructive insect pests. However, there exists a trade-off between pest resistance and lower potential yield. In the presence of uncertainty of pest severity, adopting CRISPR rice demonstrates positive environmental benefits at its optimal planting ratio, estimated based on a microeconomic model extended with environmental externalities of rice cultivation. We estimate the optimal planting ratio to be 37%, with the environmental benefit of co-planting CRISPR rice to be 560 million US dollars annually in China. The environmental benefit accounts for 4–22% of the total value of co-planting CRISPR rice in the Monte Carlo simulations. Regional heterogeneity regarding optimal planting ratio and environmental benefit is studied for 12 major rice-cultivating provinces in China. We conclude with policy implications that policymakers need to consider the vast environmental benefit of CRISPR rice adoption to have a more comprehensive view of its economic and environmental market potential, contributing to the heated debate on regulating CRISPR technology in China and worldwide.

The Use of CRISPR-Cas9 Genetic Technology in Cardiovascular Disease: A Comprehensive Review of Current Progress and Future Prospective.

Over the last century, there have been major landmark developments in the field of medicine, enabling us to control and cure various diseases on a larger scale. A few of these include the discovery of antibiotics, the development of vaccines, and the origin of organ and tissue transplants. The continued quest for innovation in microbiology and medicine has helped humankind save millions of lives and decrease morbidity at the global level.Genetic medicine has grown significantly in the last two decades and appears to be the next frontier of curative therapies for chronic diseases. One important landmark in genetic medicine is the development of CRISPR (clustered, regularly interspaced short palindromic repeats) technology. In this article, we describe the basic structure and function of the CRISPR-Cas9 system, which, simply put, consists of an RNA part and a protein. It works as a molecular scissor that can perform targeted cuts followed by repairs in and around the genes of interest to attain favorable translational outcomes. We focused on summarizing recent studies using CRISPR-Cas9 technology in diagnosing and treating cardiovascular disease. These studies are primarily experimental and limited to animal models. However, their results are promising enough to anticipate that this technology will undoubtedly be available in clinical medicine in the coming years. CRISPR-Cas9-mediated gene editing has been used to study and potentially treat congenital heart disease, hyperlipidemias, arrhythmogenic cardiomyopathies, and the prevention of ischemia-reperfusioninjury. Despite the current progress, we recognize the several challenges this technology faces, including funding for research, improving precision and reproducible results for human subjects, and establishing protocols for ethical compliance so that it is acceptable to the scientific community and the general public.

Universal crRNA Acylation Strategy for Robust Photo-Initiated One-Pot CRISPR-Cas12a Nucleic Acid Diagnostics.

Spatiotemporal regulation of clustered regularly interspaced short palindromic repeats (CRISPR) system is attractive for precise gene editing and accurate molecular diagnosis. Although many efforts have been made, versatile and efficient strategies to control CRISPR system are still desirable. Here, we proposed a universal and accessible acylation strategy to regulate the CRISPR-Cas12a system by efficient acylation of 2'-hydroxyls (2'-OH) on crRNA strand with photolabile agents (PLGs). The introduction of PLGs confers efficient suppression of crRNA function and rapid restoration of CRISPR-Cas12a reaction upon short light exposure regardless of crRNA sequences. Based on this strategy, we constructed a universal PhotO-Initiated CRISPR-Cas12a system for Robust One-pot Testing (POIROT) platform integrated with recombinase polymerase amplification (RPA), which showed two orders of magnitude more sensitive than the conventional one-step assay and comparable to the two-step assay. For clinical sample testing, POIROT achieved high-efficiency detection performance comparable to the gold-standard quantitative PCR (qPCR) in sensitivity and specificity, but faster than the qPCR method. Overall, we believe the proposed strategy will promote the development of many other universal photo-controlled CRISPR technologies for one-pot assay, and even expand applications in the fields of controllable CRISPR-based genomic editing, disease therapy, and cell imaging.

Developing a Novel Enzalutamide-Resistant Prostate Cancer Model via AR F877L Mutation in LNCaP Cells.

Prostate cancer is a leading diagnosis and major cause of cancer-related deaths in men worldwide. As a typical hormone-responsive disease, prostate cancer is commonly managed with androgen deprivation therapy (ADT) to curb its progression and potential metastasis. Unfortunately, progression to castration-resistant prostate cancer (CRPC), a notably more aggressive phase of the disease, occurs within a timeframe of 2-3 years following ADT. Enzalutamide, a recognized androgen receptor (AR) antagonist, has been employed as a standard of care for men with metastatic castration-resistant prostate cancer (mCRPC) since it was first approved in 2012, due to its ability to prolong survival. However, scientific evidence suggests that sustained treatment with AR antagonists may induce acquired AR mutations or splice variants, such as AR F877L, T878A, and H875Y, leading to drug resistance and thereby diminishing the therapeutic efficacy of these agents. Thus, the establishment of prostate cancer models incorporating these particular mutations is essential for developing new therapeutic strategies to overcome such resistance and evaluate the efficacy of next-generation AR-targeting drugs. We have developed a CRISPR (clustered regularly interspaced short palindromic repeats)-based knock-in technology to introduce an additional F877L mutation in AR into the human prostate cell line LNCaP. This article provides comprehensive descriptions of the methodologies for cellular gene editing and establishment of an in vivo model. Using these methods, we successfully identified an enzalutamide-resistant phenotype in both in vitro and in vivo models. We also assessed the efficacy of target protein degraders (TPDs), such as ARV-110 and ARV-667, in both models, and the corresponding validation data are also included here. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Generation of AR F877L-mutated LNCaP cell line using CRISPR technology Basic Protocol 2: Validation of drug resistance in AR F877L-mutated LNCaP cell line using the 2D CTG assay Support Protocol: Testing of sgRNA efficiency in HEK 293 cells Basic Protocol 3: Validation of drug resistance in AR F877L-mutated LNCaP cell line in vivo.

Advancing CRISPR technologies in reproductive biology

Clustered regularly interspaced short palindromic repeats (CRISPR) technology, a transformative tool for genetic modifications, gene therapy, and treating various genetic diseases, has recently garnered significant attention for its applications in reproductive biology. In this realm, researchers are focusing on addressing gynecological disorders, refining assisted reproductive methods, and conducting precise germ cell editing. The potential impact of CRISPR in these areas is monumental; however, several research gaps persist, demanding further investigation into the long-term effects, safety implications, and unintended consequences of its applications in reproductive biology. Refining the precision and specificity of CRISPR technology is a critical aspect that necessitates addressing challenges such as off-target effects and cytotoxicity. These considerations underscore the urgency for ongoing research efforts to ensure the efficacy and safety of CRISPR applications. In addition, the establishment of robust regulatory frameworks and oversight mechanisms specific to CRISPR in reproductive contexts is imperative to guarantee the responsible and ethical use of this powerful technology. This comprehensive review delves into the advantages of CRISPR over traditional gene editing methods, providing insights into its applications in embryos, pluripotent stem cells, and germline cells. It further explores the risks and limitations associated with CRISPR, including ethical concerns related to designer babies and eugenics. The article sheds light on the regulatory landscape governing new CRISPR applications in reproductive biology, emphasizing the continuous improvements in the efficiency and specificity of CRISPR technology. In contributing to the ongoing discourse, this review aims to inform and guide the responsible and effective application of CRISPR in the dynamic field of reproductive biology.

The Italian breakthrough in CRISPR trials for rare diseases: a focus on beta-thalassemia and sickle cell disease treatment.

The development of gene therapy and the current advantageous method of clustered regularly interspaced short palindromic repeats (CRISPRs) has allowed the implementation of several clinical trials aimed at studying the possible efficacy of gene therapy for rare diseases. Rare diseases pose a global challenge, in that their collective impact on health systems is considerable, whereas their individually rare occurrence hinders research and development of efficient therapies. Despite the low prevalence of individual rare diseases, there are more than 7,000 defined rare diseases affecting 3.5–5.9% of the global population. Rare diseases are mostly chronic and approximately 80% are caused by genetic mutation with an early-life onset. In Italy, in 2021 were recorded more than 400,000 people with rare disease. Because of its location and history, Italy has an unfortunate statistic regarding the presence and prevalence of two rare genetic diseases, namely beta-thalassemia, of which there are about 90 million carriers worldwide, 400,000 of whom are actually affected, and sickle cell disease, with about 300 million carriers and 6.5 million people affected worldwide. Advancements in genomic studies allowed Italy to join clinical trials to study effective and resolving gene therapies for BT and SCD. This study reports on the impact of rare diseases in Italy, ongoing studies, and recent achievements in BT and SCD trials using the CRISPR method and remaining hurdles in the application of CRISPR technology to rare diseases, also taking a glimpse at the newest challenges and future opportunities in the genetic treatment for rare diseases.

Nanomaterials for intelligent CRISPR-Cas tools: improving environment sustainability.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) is a desirable gene modification tool covering a wide area in various sectors of medicine, agriculture, and microbial biotechnology. The role of this incredible genetic engineering technology has been extensively investigated; however, it remains formidable with cargo choices, nonspecific delivery, and insertional mutagenesis. Various nanomaterials including lipid, polymeric, and inorganic are being used to deliver the CRISPR-Cas system. Progress in nanomaterials could potentially address these challenges by accelerating precision targeting, cost-effectiveness, and one-step delivery. In this review, we highlighted the advances in nanotechnology and nanomaterials as smart delivery systems for CRISPR-Cas so as to ameliorate applications for environmental remediation including biomedical research and healthcare, strategies for mitigating antimicrobial resistance, and to be used as nanofertilizers for enhancing crop growth, and reducing the environmental impact of traditional fertilizers. The timely co-evolution of nanotechnology and CRISPR technologies has contributed to smart novel nanostructure hybrids for improving the onerous tasks of environmental remediation and biological sustainability.

Challenges with Newly Approved CRISPR Gene Technologies

CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) technology is a powerful gene-editing tool that allows for precise modification of DNA. While CRISPR has shown immense potential in laboratory and preclinical studies, the development of CRISPR-based therapies for human use is still in the early stages. However, on December 08, the Food and Drug Administration (FDA) of the United States of America approved two milestone treatments, Casgevy and Lyfgenia, representing the first cell-based gene therapies for the treatment of sickle cell disease (SCD) in patients 12 years and older. Several CRISPR-Cas9 therapies that work on the same principle are in clinical trials as treatments for a range of diseases. However, there are still significant concerns with the use of this new technology. One major concern with CRISPR technology is the potential for off-target effects, where unintended changes may occur in locations other than the target gene.  The introduction of CRISPR components into the body may also trigger an immune response. Several laboratories are investigating ways to mitigate immune reactions and improve the delivery of CRISPR components. The method of delivering CRISPR components into cells is also important for safety. Further research into different delivery methods, such as viral vectors or nanoparticles, would help optimize efficiency while minimizing potential side effects. The long-term effects of CRISPR-based interventions are an area of ongoing research. Understanding the stability of CRISPR-induced changes over time and potential consequences is crucial for assessing safety. Ethical considerations are important to discussions about CRISPR use, particularly when it comes to editing the human germline (sperm, eggs, embryos), as changes made in these cells would be heritable. The potential for unintended consequences and ethical implications necessitates careful consideration. It is important to consider that although CRISPR holds great promise, the technology is relatively young, and research is ongoing to address safety concerns. The scientific community and regulatory bodies are actively working to ensure that CRISPR technologies are developed and implemented responsibly. Continuous advancements, ongoing research, and open communication will be essential to addressing safety challenges associated with CRISPR.

Advancing Crop Improvement Through CRISPR Technology in Precision Agriculture Trends-A Review

The advent of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) technology has ushered in a new era in agricultural biotechnology, offering unprecedented opportunities for targeted genome editing and crop improvement. This review article presents a comprehensive examination of the advancements, applications, challenges, and future prospects of CRISPR technology within the context of precision agriculture. The integration of CRISPR with precision agriculture technologies signifies a major shift towards more efficient and sustainable farming practices, emphasizing the precise modification of crops to enhance yield, disease resistance, and environmental stress tolerance. The historical backdrop of agricultural biotechnology and the evolution of precision agriculture set the stage for understanding the transformative impact of CRISPR technology. CRISPR's superiority over traditional breeding and genetic modification techniques lies in its precision, speed, and cost-effectiveness. Detailed case studies of CRISPR-modified crops, such as disease-resistant wheat, drought-tolerant rice, and nutrient-efficient maize, highlight the technology's practical implications. These modifications not only enhance crop performance but also contribute to ecological sustainability and increased farmer income, demonstrating CRISPR's significant role in addressing global food security challenges. The application of CRISPR in agriculture is not without challenges. Regulatory hurdles, public perception, technical limitations, and ethical considerations present substantial obstacles to the widespread adoption of CRISPR-modified crops. The review addresses these challenges, offering insights into the complex interplay between technological innovation and societal acceptance. Further explores potential developments in CRISPR technology, including next-generation genome editing tools and the integration of synthetic biology. It underscores the importance of interdisciplinary collaborations and adaptive policy frameworks to navigate the evolving technological and regulatory landscapes. The future of CRISPR in precision agriculture promises not only enhanced crop varieties but also a paradigm shift towards more data-driven, customized, and environmentally conscious farming practices. This review concludes that CRISPR technology, despite its challenges, holds immense promise for revolutionizing agriculture. Its continued development and responsible implementation are key to realizing its full potential in contributing to a sustainable and secure agricultural future.

Applications of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) as a Genetic Scalpel for the Treatment of Cancer: A Translational Narrative Review.

Cancer remains a global health challenge with high prevalence and mortality rates, imposing significant financial and emotional burdens on affected families. However, hope lies in genetic manipulation, with a focus on innovative approaches to combat genetically linked cancers. Clustered regularly interspaced short palindromic repeats (CRISPR), an adaptive immune system found in various bacteria and archaea, hold immense potential. We searched articles on PubMed Central, Medline, and PubMed databasesindexed journals. The keywords from the research topic, i.e., "CRISPR AND genetic therapy," were searched, and we found 3397 articles. Following this, we explored the medical subject headings (MeSH) glossary and created a search strategy "Clustered Regularly Interspaced Short Palindromic Repeats"[Mesh] AND "Genetic Therapy"[Majr] and after applying a variety of filters we included 30 studies in our review. CRISPR consists of unique spacers and CRISPR-associated (Cas) genes, operating through acquisition, CRISPR ribonucleic acid (crRNA) biogenesis, and target interference phases. The type II CRISPR-Cas9 system is a well-researched avenue for gene editing, with Cas9 cleaving specific genomic regions and initiating deoxyribonucleic acid (DNA) repair mechanisms. Cancer results from genetic alterations, leading to tumor development with properties like metastasis. CRISPR/Cas9 offers precise genome editing to inhibit tumor formation by removing specific genomic sequences and promoting DNA repair. Challenges in CRISPR's use for cancer therapy, including delivery methods, cell adaptation, and ethical concerns, are recognized. Beyond cancer, CRISPR finds diverse applications in infectious diseases and non-cancerous conditions, signifying its transformative potential in modern medicine. CRISPR technology represents a groundbreaking frontier in cancer therapy and beyond, offering insights into genetic editing, disease mechanisms, and potential cures. Despite challenges and ethical considerations, precise genome editing promises improved cancer treatments and innovative medical interventions in the future.

Global research trends in CRISPR-related technologies associated with extracellular vesicles from 2015 to 2022: a bibliometric, dynamic, and visualized study

PurposeThis study aims to explore the emerging trends, dynamic development, and research hotspots of clustered regularly interspaced short palindromic repeats (CRISPR) technology associated with extracellular vesicles during the past 7 years and demonstrate them by visualization.MethodsA total of 219 records related to CRISPR technology associated with extracellular vesicles from 2015 to 2022 in the Web of Science Core Collection (WoSCC) database were collected. R language, VOSviewer, CiteSpace, and GraphpadPrism software packages were used to analyze the history of this research, the general characteristics of the literature, and keywords. Finally, the hotspots and latest trends in CRISPR technology associated with extracellular vesicles are predicted.ResultsA total of 219 articles were collected for this study. The production of publications about CRISPR technology associated with extracellular vesicles has increased annually. Researchers from China, the USA, and Germany made the most important contributions to this trend, while RLUK Research Libraries UK offers the largest amount of literature in this field. Shenzhen University, Nanjing Medicine University, and Peking University exhibited the closest cooperation. Additionally, active topics burst during different periods, as identified according to 317 keywords belonging to 39 disciplines. Keywords were clustered into seven research subareas, namely exosome, nanovesicles, DNA, gene editing, gene therapy, cancer therapy, and endometrial stromal cells. The alluvial map of keywords reveals that the most enduring concepts are gene therapy, nanovesicles, etc., while the emerging keywords are genome, protein delivery, plasma, etc.ConclusionsWe reviewed 219 previous publications and conducted the first bibliometric study of CRISPR technology related to extracellular vesicles from 2015 to 2022. This comprehensive summary constructed a knowledge map and demonstrates the trends in this area. The current trends and potential hotpots for this topic are also identified, which will be a great help for researchers in the future.

Harnessing CRISPR/Cas9 for Enhanced Disease Resistance in Hot Peppers: A Comparative Study on CaMLO2-Gene-Editing Efficiency across Six Cultivars

The Capsicum annuum Mildew Locus O (CaMLO2) gene is vital for plant defense responses against fungal pathogens like powdery mildew, a significant threat to greenhouse pepper crops. Recent advancements in genome editing, particularly using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9, have unlocked unprecedented opportunities for modifying disease-resistant genes and improving crop characteristics. However, the application of CRISPR technology in pepper cultivars has been limited, and the regeneration process remains challenging. This study addresses these limitations by investigating the feasibility of using the validated CaMLO2 genetic scissors system in six commercial hot pepper cultivars. We assessed the gene-editing efficiency of the previously reported high-efficiency Cas9/CaMLO2single-guide RNA (sgRNA)1-ribonucleoprotein (RNP) and the low-efficiency Cas9/CaMLO2sgRNA2-RNP systems by extending their application from the bell pepper ‘Dempsey’ and the hot pepper ‘CM334’ to six commercial hot pepper cultivars. Across the six cultivars, CaMLO2sgRNA1 demonstrated an editing efficiency ranging from 6.3 to 17.7%, whereas CaMLO2sgRNA2 exhibited no editing efficiency, highlighting the superior efficacy of sgRNA1. These findings indicate the potential of utilizing the verified Cas9/CaMLO2sgRNA1-RNP system to achieve efficient gene editing at the CaMLO2 locus in different Capsicum annuum cultivars regardless of their cultivar genotypes. This study provides an efficacious genome-editing tool for developing improved pepper cultivars with CaMLO2-mediated enhanced disease resistance.

Advancements in the research of combining MRI and CRISPR technology for the treatment of breast cancer

Cancer remains a significant contributor to global mortality, posing a considerable public health challenge. Among the various types of cancer, breast cancer stands out as one of the most prevalent among women worldwide. Ongoing efforts are dedicated to the development of innovative treatments with minimal side effects. A groundbreaking genome technique for editing, CRISPR-associated protein-9 (Cas9), also known as clustered regularly interspaced short palindromic repeat (CRISPR), has become an effective instrument for genetic modification. This technology has revolutionized genome editing across diverse biological fields, including its applications in genome therapy and tumor research. With ongoing advancements in sensitivity, specificity, and diagnostic accuracy, magnetic resonance imaging (MRI) has gained increasing recognition for the detection of breast cancer. MRIs capability to produce high-resolution images plays a pivotal role in aiding healthcare professionals in precise localization of breast masses, tumors, and other abnormalities. Additionally, it facilitates the assessment of parameters like tumor size, extent, and lymph node involvement, contributing to the formulation of more accurate treatment strategies. Here is a review summary of the combined utilization of MRI and CRISPR technology in breast cancer treatment. It explores the synergy between these two technologies, discussing their advantages and disadvantages and their potential to enhance diagnostic and therapeutic outcomes.

An amplification-free CRISPR-SERS biosensor for specific, sensitive and rapid detection of Salmonella Typhimurium in poultry

CRISPR (Clustered regularly interspaced short palindromic repeats) has been a cutting-edge biotechnology in both gene editing and biosensing due to its desirable features, such as high base resolution and set-and-forget operation. However, as a nucleic acid detection method, an DNA/RNA amplification procedure is still inevitable in CRISPR biosensors. In this study, a highly-ordered nanopore array SERS substrate, coated with silver nanoparticles, was fabricated and utilized as a platform for the specific detection of Salmonella Typhimurium (S. Typhimurium) without the time-consuming DNA amplification step. The smart sensing strategy was established by introducing an ssDNA labeled with a Raman active molecule as a signal report probe to show the changes during target identification and detection. The CRISPR-SERS biosensor was developed by combining the ultra-sensitive sensing mechanism of CRISPR technology with the SERS analytical method, aiming at detecting the invasin A gene (invA) of S. Typhimurium. The sensing materials and operation conditions were optimized to maximize the output signal and minimize the cost. The results showed that the proposed biosensor could specifically detect S. Typhimurium with a limit of detection (LOD) of 110 CFU/mL in poultry samples within 2 h. This CRISPR-SERS sensing strategy could provide an alternative way for pathogen detection in food safety monitoring.

The Cutting-edge of CRISPR for Cancer Treatment and its Future Prospects.

CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is a versatile technology that allows precise modification of genes. One of its most promising applications is in cancer treatment. By targeting and editing specific genes involved in cancer development and progression, CRISPR has the potential to become a powerful tool in the fight against cancer. This review aims to assess the recent progress in CRISPR technology for cancer research and to examine the obstacles and potential strategies to address them. The two most commonly used CRISPR systems for gene editing are CRISPR/Cas9 and CRISPR/Cas12a. CRISPR/Cas9 employs different repairing systems, including homologous recombination (HR) and nonhomologous end joining (NHEJ), to introduce precise modifications to the target genes. However, off-target effects and low editing efficiency are some of the main challenges associated with this technology. To overcome these issues, researchers are exploring new delivery methods and developing CRISPR/Cas systems with improved specificity. Moreover, there are ethical concerns surrounding using CRISPR in gene editing, including the potential for unintended consequences and the creation of genetically modified organisms. It is important to address these issues through rigorous testing and strict regulations. Despite these challenges, the potential benefits of CRISPR in cancer therapy cannot be overlooked. By introducing precise modifications to cancer cells, CRISPR could offer a targeted and effective treatment option for patients with different types of cancer. Further investigation and development of CRISPR technology are necessary to overcome the existing challenges and harness its full potential in cancer therapy.

CRISPR gene editing to improve crop resistance to parasitic plants.

Parasitic plants pose a significant threat to global agriculture, causing substantial crop losses and hampering food security. In recent years, CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) gene-editing technology has emerged as a promising tool for developing resistance against various plant pathogens. Its application in combating parasitic plants, however, remains largely unexplored. This review aims to summarise current knowledge and research gaps in utilising CRISPR to develop resistance against parasitic plants. First, we outline recent improvements in CRISPR gene editing tools, and what has been used to combat various plant pathogens. To realise the immense potential of CRISPR, a greater understanding of the genetic basis underlying parasitic plant-host interactions is critical to identify suitable target genes for modification. Therefore, we discuss the intricate interactions between parasitic plants and their hosts, highlighting essential genes and molecular mechanisms involved in defence response and multilayer resistance. These include host resistance responses directly repressing parasitic plant germination or growth and indirectly influencing parasitic plant development via manipulating environmental factors. Finally, we evaluate CRISPR-mediated effectiveness and long-term implications for host resistance and crop improvement, including inducible resistance response and tissue-specific activity. In conclusion, this review highlights the challenges and opportunities CRISPR technology provides to combat parasitic plants and provides insights for future research directions to safeguard global agricultural productivity.

Applications of CRISPR-Cas9 for advancing precision medicine in oncology: from target discovery to disease modeling.

The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) (CRISPR/Cas9) system is a powerful tool that enables precise and efficient gene manipulation. In a relatively short time, CRISPR has risen to become the preferred gene-editing system due to its high efficiency, simplicity, and programmability at low costs. Furthermore, in the recent years, the CRISPR toolkit has been rapidly expanding, and the emerging advancements have shown tremendous potential in uncovering molecular mechanisms and new therapeutic strategies for human diseases. In this review, we provide our perspectives on the recent advancements in CRISPR technology and its impact on precision medicine, ranging from target identification, disease modeling, and diagnostics. We also discuss the impact of novel approaches such as epigenome, base, and prime editing on preclinical cancer drug discovery.

Evolving Therapeutic Options for the Treatment of Duchenne Muscular Dystrophy

Duchenne muscular dystrophy (DMD) is the most common childhood form of muscular dystrophy. It is caused by mutations in the DMD gene, leading to reduced or absent expression of the dystrophin protein. Clinically, this results in loss of ambulation, cardiomyopathy, respiratory failure, and eventually death. In the past decades, the use of corticosteroids has slowed down the disease progression. More recently, the development of genetically mediated therapies has emerged as the most promising treatment for DMD. These strategies include exon skipping with antisense oligonucleotides, gene replacement therapy with adeno-associated virus, and gene editing with CRISPR (clustered regularly interspaced short palindromic repeats) technology. In this review, we highlight the most up-to-date therapeutic progresses in the field, with emphasis on past and recent experiences, as well as the latest clinical results of DMD micro-dystrophin gene therapy. Additionally, we discuss the lessons learned along the way and the challenges encountered, all of which have helped advance the field, with the potential to finally alleviate such a devastating disease.

Applications of CRISPR Technology to Breast Cancer and Triple Negative Breast Cancer Research.

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology has transformed oncology research in many ways. Breast cancer is the most prevalent malignancy globally and triple negative breast cancer (TNBC) is one of the most aggressive subtypes with numerous challenges still to be faced. In this work, we have explained what CRISPR consists of and listed its applications in breast cancer while focusing on TNBC research. These are disease modelling, the search for novel genes involved in tumour progression, sensitivity to drugs and immunotherapy response, tumour fitness, diagnosis, and treatment. Additionally, we have listed the current delivery methods employed for the delivery of CRISPR systems in vivo. Lastly, we have highlighted the limitations that CRISPR technology is subject to and the future directions that we envisage. Overall, we have provided a round summary of the aspects concerning CRISPR in breast cancer/TNBC research.