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Challenges and advances of CRISPR-Cas9 genome editing in therapeutics.
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CRISPR-Cas12a System for Biosensing and Gene Regulation.
Clustered regularly interspaced short palindromic repeats (CRISPR) is a promising technology in the biological world. As one of the CRISPR-associated (Cas) proteins, Cas12a is an RNA-guided nuclease in the type V CRISPR-Cas system, which has been a robust tool for gene editing. In addition, due to the discovery of target-binding-induced indiscriminate single-stranded DNase activity of Cas12a, CRISPR-Cas12a also exhibits great promise in biosensing. This minireview not only gives a brief introduction to the mechanism of CRISPR-Cas12a but also highlights the recent developments and applications in biosensing and gene regulation. Finally, future prospects of the CRISPR-Cas12a system are also discussed. We expect this minireview will inspire innovative work on the CRISPR-Cas12a system by making full use of its features and advantages.
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CRISPR/Cas in Arabidopsis: overcoming challenges to accelerate improvements in crop photosynthetic efficiencies.
The rapid and widespread adoption of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas technologies has allowed genetic editing in plants to enter a revolutionary new era. In this mini review, we highlight the current CRISPR/Cas tools available in plants and the use of Arabidopsis thaliana as a model to guide future improvements in crop yields, such as enhancing photosynthetic potential. We also outline the current socio-political landscape for CRISPR/Cas research and highlight the growing need for governments to better facilitate research into plant genetic-editing technologies.
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Panacea in progress: CRISPR and the future of its biological research introduction.
The elucidation of the CRISPR (clustered, regularly interspaced, short palindromic repeats) adaptive immune system endogenous to most microbial life has culminated in progress in a diversity of scientific disciplines. The concurrently promising and eccentric nature of its theoretically plausible applications has wrought enthusiasm in the research community globally, potentiating advancements in human and animal health, ecological stability, and economic wellbeing, that would hitherto be considered the unattainable fancies of a futurist. It may be supposed that the tomes of science fiction are the true books of prophecy. Here, we narrate the scientific dialogue regarding CRISPR/Cas biotechnologies, from the happenstantial initial observation of the locus to the litany of intriguing contemporary endeavors. We discuss the mechanistic underpinnings in detail, and the corpulent body of literature on CRISPR-based biotech is digested into a germane and informative review. CRISPR applications such as microbiome engineering in order to enhance the human immune system beyond the fortitude of the wild type, bacterial genome editing in industrial and medical aspects, conquering antibiotic resistance, the development of novel antimicrobial techniques, the harvesting of solventogenic microbes, the development of antifungal therapies, and investigation of the genetic properties of fungi, are here represented, and the authors posit unconventional, and at times gainfully tangential, thoughts and concepts in order to encourage a reflective disposition towards this sophisticated device of nature: a panacea in progress, such that the most impassive and technical writing still carries the ring of poetry.
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CRISPR: a precise genome editing strategy for the treatment of hepatocellular carcinoma.
INTRODUCTION: The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) gene-editing tool provides novel therapeutic alternatives by promoting the gene alteration in adaptive T cells or malignant cells to combat Hepatocellular Carcinoma (HCC). More successful cancer treatments are now possible due to the capacity of precisely locating and modifying particular genetic abnormalities that promote malignancy growth and metastasis. AREAS COVERED: In this review, we address ongoing clinical trials, the possible similarities between CRISPR-based cancer treatments and current therapeutic choices, and how CRISPR technology can improve treatment outcomes for HCC while using the latest safety measures. Additionally, this analysis sheds light on the existing obstacles and potential future possibilities of applying CRISPR technology to the management of HCC, with a final objective of enhancing patient results and completely changing the field of HCC therapies. EXPERT OPINION: The urgent need for innovative therapies is underscored by the poor prognosis associated with severe hepatocellular carcinoma, despite recent advancements in clinical therapies. Through a special emphasis on invivo cancer cell targeting along with the generation of chimeric antigen receptor (CAR) T cells, including T cell receptor (TCR) T cells, this review analyses the uses of CRISPR methods in the therapy of HCC.
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Genome-Editing Technologies: Principles and Applications.
Targeted nucleases have provided researchers with the ability to manipulate virtually any genomic sequence, enabling the facile creation of isogenic cell lines and animal models for the study of human disease, and promoting exciting new possibilities for human gene therapy. Here we review three foundational technologies-clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9), transcription activator-like effector nucleases (TALENs), and zinc-finger nucleases (ZFNs). We discuss the engineering advances that facilitated their development and highlight several achievements in genome engineering that were made possible by these tools. We also consider artificial transcription factors, illustrating how this technology can complement targeted nucleases for synthetic biology and gene therapy.
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Genome-wide CRISPR screening identifies cellular factors controlling nonviral genome editing efficiency.
To systematically map cellular factors constraining nonviral genome editing, influencing uptake and intracellular trafficking, we develop a genome-wide CRISPR screening platform linking perturbation of 19,114 genes to editing outcomes in human cells. We identify six negative regulators of delivery whose depletion increases editing efficiency by up to six-fold across diverse payloads, loci, and cell types. We test the top two factors, GJB2 and BET1L, in two distinct human models: correction of a pathogenic adenine base mutation in KCNJ13 and introduction of a cytosine base mutation in the GABAA receptor gene. Depletion of either improves base-editing outcomes by 6-fold, potentially through effects on delivery. In a patient-derived model of retinal channelopathy, knockdown of either gene improves lipid nanoparticle base editing efficiency by over 3.5-fold. This enables functional restoration of Kir7.1 ion channels in a subset of edited cells, highlighting cellular barriers as actionable targets to enhance the potency of genetic therapies.
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CRISPR Screening "Big Data" Informs Novel Therapeutic Solutions.
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CRISPR/dCas9 for hepatic fibrosis therapy: implications and challenges.
Hepatic fibrosis is a pathological reaction of tissue damage and repair caused by various pathogenic factors acting on liver. At present, there is no effective anti-fibrotic specific therapy. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (dCas9) system is a new generation of gene editing technology. The CRISPR/dCas9 system provides a platform for studying site-specific transcriptional regulation, which has high efficiency in gene transcriptional activation for achieving robust. This system holds promise for hepatic fibrosis therapy via acting on liver fibrosis effector cells. However, there are some challenges associated with this novel technology, such as large structural variants at on-target, off-target sites, and targeted delivery efficiency. In this review, we present the potential implications and describe the challenges of CRISPR/dCas9 system that might be encountered in hepatic fibrosis therapy.
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Guiding Lights in Genome Editing for Inherited Retinal Disorders: Implications for Gene and Cell Therapy.
Inherited retinal dystrophies (IRDs) are a leading cause of visual impairment in the developing world. These conditions present an irreversible dysfunction or loss of neural retinal cells, which significantly impacts quality of life. Due to the anatomical accessibility and immunoprivileged status of the eye, ophthalmological research has been at the forefront of innovative and advanced gene- and cell-based therapies, both of which represent great potential as therapeutic treatments for IRD patients. However, due to a genetic and clinical heterogeneity, certain IRDs are not candidates for these approaches. New advances in the field of genome editing using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas) have provided an accurate and efficient way to edit the human genome and represent an appealing alternative for treating IRDs. We provide a brief update on current gene augmentation therapies for retinal dystrophies. Furthermore, we discuss recent advances in the field of genome editing and stem cell technologies, which together enable precise and personalized therapies for patients. Lastly, we highlight current technological limitations and barriers that need to be overcome before this technology can become a viable treatment option for patients.
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