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CRISPR activation screens: navigating technologies and applications.
Clustered regularly interspaced short palindromic repeats (CRISPR) activation (CRISPRa) has become an integral part of the molecular biology toolkit. CRISPRa genetic screens are an exciting high-throughput means of identifying genes the upregulation of which is sufficient to elicit a given phenotype. Activation machinery is continually under development to achieve greater, more robust, and more consistent activation. In this review, we offer a succinct technological overview of available CRISPRa architectures and a comprehensive summary of pooled CRISPRa screens. Furthermore, we discuss contemporary applications of CRISPRa across broad fields of research, with the aim of presenting a view of exciting emerging applications for CRISPRa screening.
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CasX: a new and small CRISPR gene-editing protein.
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Therapeutic Genome Editing by CRISPR/Cas9-Mediated Strategy to Cure Genetic Disorders in Humans: Guide for Molecular Surgeons.
Genetic engineering scientists have shown a wide increase especially on therapeutic promise which has gained considerable awareness. This goes back to a variety of tools including the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system. This technique has shown an efficient flourish in genome editing and treatment of genetic diseases. Due to its specificity and ease of use it has been widely adopted for a variety of applications involving human diseases, treatment, therapeutics in cancer and biotechnology. Albeit studies have shown that CRISPR/Cas9 technique is more proficient, particular and manipulable than past eras of genetic modulation instruments. Additionally, it can be further enhanced by the application of different developments raising its general productivity in a higher recurrence of genome modifications and decreasing its off-target impacts. Here in this review this technique will be introduced with discussing its relation with the genome engineering and gene editing especially in the therapeutic area. Furthermore, the specificity of the application is explained and the contribution of Induced Human Pluripotent Stem Cells (iPSCs) in gene manipulation and how CRISPR technology expanded its application is discussed. Lastly, we describe the novel effective applications of the CRISPR/Cas9 technology and propose some enhancements that might improve the technique with highlighting the future approach and remarks.
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Vaccinia Virus Genome Editing Using CRISPR.
CRISPR/Cas9, an RNA-guided targeted genome editing system, can make precise, targeted modifications to the genome in living cells. Here we describe how this method can be used to efficiently edit the vaccinia virus genome enabling the insertion of transgene(s) specifically into a targeted site.
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CRISPR Gets Its Origin Story.
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Progress and challenges in CRISPR/Cas applications in microalgae.
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technologies have emerged as powerful tools for precise genome editing, leading to a revolution in genetic research and biotechnology across diverse organisms including microalgae. Since the 1950s, microalgal production has evolved from initial cultivation under controlled conditions to advanced metabolic engineering to meet industrial demands. However, effective genetic modification in microalgae has faced significant challenges, including issues with transformation efficiency, limited target selection, and genetic differences between species, as interspecies genetic variation limits the use of genetic tools from one species to another. This review summarized recent advancements in CRISPR systems applied to microalgae, with a focus on improving gene editing precision and efficiency, while addressing organism-specific challenges. We also discuss notable successes in utilizing the class 2 CRISPR-associated (Cas) proteins, including Cas9 and Cas12a, as well as emerging CRISPR-based approaches tailored to overcome microalgal cellular barriers. Additionally, we propose future perspectives for utilizing CRISPR/Cas strategies in microalgal biotechnology.
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The CRISPR Patent Ruling and Implications for Medicine.
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Exogenous gene integration mediated by genome editing technologies in zebrafish.
Genome editing technologies, such as transcription activator-like effector nuclease (TALEN) and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) systems, can induce DNA double-strand breaks (DSBs) at the targeted genomic locus, leading to frameshift-mediated gene disruption in the process of DSB repair. Recently, the technology-induced DSBs followed by DSB repairs are applied to integrate exogenous genes into the targeted genomic locus in various model organisms. In addition to a conventional knock-in technology mediated by homology-directed repair (HDR), novel knock-in technologies using refined donor vectors have also been developed with the genome editing technologies based on other DSB repair mechanisms, including non-homologous end joining (NHEJ) and microhomology-mediated end joining (MMEJ). Therefore, the improved knock-in technologies would contribute to freely modify the genome of model organisms.
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CRISPR in 3D: Innovations in Disease Modelling and Personalized Medicine.
The CRISPR-Cas system of genetic engineering has had a significant impact on science and society since its advent in 2013. CRISPR integration with 3D culture systems such as organ-on-a-chips, as well as fast emerging commercial technologies, has encouraged translation of more complex pathophysiological modelling and personalized medicine.
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CRISPR Technology and Its Emerging Applications.
The discovery and iteration of clustered regularly interspaced short palindromic repeats (CRISPR) systems have revolutionized genome editing due to their remarkable efficiency and easy programmability, enabling precise manipulation of genomic elements. Owing to these unique advantages, CRISPR technology has the transformative potential to elucidate biological mechanisms and develop clinical treatments. This review provides a comprehensive overview of the development and applications of CRISPR technology. After describing the three primary CRISPR-Cas systems - CRISPR-associated protein 9 (Cas9) and Cas12a targeting DNA, and Cas13 targeting RNA - which serve as the cornerstone for technological advancements, we describe a series of novel CRISPR-Cas systems that offer new avenues for research, and then explore the applications of CRISPR technology in large-scale genetic screening, lineage tracing, genetic diagnosis, and gene therapy. As this technology evolves, it holds significant promise for studying gene functions and treating human diseases in the near future.
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