CRISPR-Cas systems: ushering in the new genome editing era (original) (raw)
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Gene, 2020
Since the discovery of the double helix and the introduction of genetic engineering, the possibility to develop new strategies to manipulate the genome has fascinated scientists around the world. Currently scientists have the knowledge and ability to edit the genomes. Several methodologies of gene editing have been established, all of them working like "scissor", creating double strand breaks at specific spots. The introduction of a new technology, which was adapted from the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas bacterial immune system, has revolutionized the genetic therapy field, as it allows a much more precise editing of gene than the previously described tools and, therefore, to prevent and treat disease in humans. This review aims to revisit the genome editing history that led to the rediscovery of the CRISPR/Cas technology and to explore the technical aspects, applications and perspectives of this fascinating, powerful, precise, simpler and cheaper technology in different fields. 1. The history of the genome editing tools
https://ijshr.com/IJSHR\_Vol.6\_Issue.3\_July2021/IJSHR-Abstract.044.html, 2021
In a way akin to how technological advancements involving molecular and biological science rapidly developed during the last decade, CRISPR, a newly found genomic modifying technique, quickly gained the attention of the scientific community. This paper aims to provide a fundamental understanding of CRISPR technology by reviewing articles from several journals, consisting of general information about CRISPR technology, process, advantages, limitations, and comparisons with other technologies. This recent technology drastically altered the boundaries of genetic engineering-most notably due to its outstanding flexibility of gRNA modification, as demonstrated by the work of the two Nobel Prize award-winning scientists Emmanuelle Charpentier and Jennifer A. Doudna. From the journals that had been reviewed, this paper presents that the CRISPR-Cas9 is one of the most efficient tools for genetic engineering in our modern world because of its incredible potential to perform genetic material modification in a wide range of patients with greater efficiency, versatility, and accuracy than before, all the while being more cost-effective. Additionally, since significant research and newfound knowledge related to genetic science was made possible from the discovery of this CRISPR technology, the possibility of CRISPR-Cas9 treatment in patients-particularly to combat congenital genetic diseases-would become a horizon that is within the reach of humanity's hands.
International Journal of Molecular Sciences, 2021
According to Darwin’s theory, endless evolution leads to a revolution. One such example is the Clustered Regularly Interspaced Palindromic Repeats (CRISPR)–Cas system, an adaptive immunity system in most archaea and many bacteria. Gene editing technology possesses a crucial potential to dramatically impact miscellaneous areas of life, and CRISPR–Cas represents the most suitable strategy. The system has ignited a revolution in the field of genetic engineering. The ease, precision, affordability of this system is akin to a Midas touch for researchers editing genomes. Undoubtedly, the applications of this system are endless. The CRISPR–Cas system is extensively employed in the treatment of infectious and genetic diseases, in metabolic disorders, in curing cancer, in developing sustainable methods for fuel production and chemicals, in improving the quality and quantity of food crops, and thus in catering to global food demands. Future applications of CRISPR–Cas will provide benefits for...
Current Applied Science and Technology, 2022
The clustered regularly interspaced short palindromic repeat paired associated protein 9 (CRISPR-Cas9) is a site-specific genome editing tool that enables scientists to edit or introduce genetic mutation at will. CRISPR-Cas9 consists of two essential key players; a programmable RNA called single guide RNA (sgRNA) and the Cas9 protein which functions as a molecular scissors that does the cutting. Since its discovery, CRISPR-Cas9 has received vast attention due to its simplicity, convenience, and superior precision of use. Its application extends into various fields including the health sciences where it has been used to enhance the understanding of pathogenesis and help in therapeutic intervention. Despite the promising potentials and applications of CRISPR-Cas9, there are several aspects that need to be addressed including the method of delivery, off-target cutting and ethical issues in human germline modification. The purposes of this review are to perform a comprehensive literature search of publications on the CRISPR-Cas9 system and to highlight potential applications of CRISPR-Cas9 in the field of medical sciences. In this present review, we discuss the background of CRISPR-Cas9, its mechanisms of genome modification and its applications in the medical field including its use in the study of animal model production, genetics, multifactorial and complex diseases. In addition, we also discuss the limitations associated with CRISPR-Cas9 application. CRISPR-Cas9 has accelerated medical studies and facillitate the collection of vast amounts of information. However, its limitations should be further studied in order to reap its greatest benefits.
Genome editing with the CRISPR‐Cas system: an art, ethics and global regulatory perspective
Plant Biotechnology Journal, 2020
Over the last three decades, the development of new genome editing techniques, such as ODM, TALENs, ZFNs and the CRISPR-Cas system, has led to significant progress in the field of plant and animal breeding. The CRISPR-Cas system is the most versatile genome editing tool discovered in the history of molecular biology because it can be used to alter diverse genomes (e.g. genomes from both plants and animals) including human genomes with unprecedented ease, accuracy and high efficiency. The recent development and scope of CRISPR-Cas system have raised new regulatory challenges around the world due to moral, ethical, safety and technical concerns associated with its applications in pre-clinical and clinical research, biomedicine and agriculture. Here, we review the art, applications and potential risks of CRISPR-Cas system in genome editing. We also highlight the patent and ethical issues of this technology along with regulatory frameworks established by various nations to regulate CRISPR-Cas-modified organisms/products. The art of CRISPR-Cas system Genome editing has revolutionized DNA manipulation in eukaryotes enabling the precise mutagenesis of single base pairs, the introduction of insertions and/or deletions (indels), DNA fragment substitution and the nucleotide base conversion (Carroll, 2017; Naso and Petrova, 2019). The most frequent genome editing systems involve ODM (oligonucleotide-directed mutagenesis; Sauer et al., 2016), TALENs (transcription activator-like effector nucleases), ZFNs (zinc finger nucleases) and CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPRassociated proteins; Carroll, 2014; Petolino, 2015). TALENs and ZFNs are still in use for research in various agricultural and medical arenas. However, the applications of these technologies are limited because of the transfection inefficiency, design complexity and limitations on multiplexed mutations (Doudna and Charpentier, 2014). In contrast, the most powerful and game-changing technique, the CRISPR-Cas system, is currently the dominant genome editing technology in research laboratories around the world due to its efficiency, precision and broad application range (Ledford, 2015). The CRISPR-Cas system was discovered as a form of adaptive immunity system in bacteria and archaea to defend against invading viral, plasmid and/or phage DNA (Albitar et al., 2018; Ishino et al., 1987; Sovov a et al., 2016). CRISPR-Cas system is typically comprised of two parts: (i) Cas proteins-involved in protection and the acquisition of invading nucleotides, and (ii) the CRISPR array-which consists of conserved recurrent domains known as direct repeats and imbedded variable sequences with same length known as spacers (Figure 1). The CRISPR array helps
Application of CRISPR-Cas9 genome editing technology in various fields: A review
narraj, 2023
CRISPR-Cas9 has emerged as a revolutionary tool that enables precise and efficient modifications of the genetic material. This review provides a comprehensive overview of CRISPR-Cas9 technology and its applications in genome editing. We begin by describing the fundamental principles of CRISPR-Cas9 technology, explaining how the system utilizes a single guide RNA (sgRNA) to direct the Cas9 nuclease to specific DNA sequences in the genome, resulting in targeted double-stranded breaks. In this review, we provide in-depth explorations of CRISPR-Cas9 technology and its applications in agriculture, medicine, environmental sciences, fisheries, nanotechnology, bioinformatics, and biotechnology. We also highlight its potential, ongoing research, and the ethical considerations and controversies surrounding its use. This review might contribute to the understanding of CRISPR-Cas9 technology and its implications in various fields, paving the way for future developments and responsible applications of this transformative technology.
CRISPR-Cas technology a new era in genomic engineering
Biotechnology Reports, 2022
The CRISPR-Cas systems have offered a flexible, easy-to-use platform to precisely modify and control the genomes of organisms in various fields, ranging from agricultural biotechnology to therapeutics. This system is extensively used in the study of infectious, progressive, and life-threatening genetic diseases for the improvement of quality and quantity of major crops and in the development of sustainable methods for the generation of biofuels. As CRISPR-Cas technology continues to evolve, it is becoming more controllable and precise with the addition of molecular regulators, which will provide benefits for everyone and save many lives. Studies on the constant growth of CRISPR technology are important due to its rapid development. In this paper, we present the current applications and progress of CRISPR-Cas genome editing systems in several fields of research, we further highlight the applications of anti-CRISPR molecules to regulate CRISPR-Cas gene editing systems, and we discuss ethical considerations in CRISPR-Cas applications.
CRISPR/Cas9 System: A Breakthrough in Genome Editing
Molecular Biology
Clustered Regularly Interspaced Short Palindromic Repeats is a new and advance gene editing tool using specific nuclease enzyme for specific cleavage. It is the most efficient technique, commonly used for many purposes like gene therapy, production of desired plants and transgenic animals. But it has some limitations like off-target issue and this issue can be minimized by the production of specific sequence of guide RNA. In the future, it can be used for the treatment of many human genetic diseases.
Advances In Research On Genome Editing Crispr-Cas9 Technology
Journal of Ayub Medical College, Abbottabad : JAMC, 2019
BACKGROUND The current era of genome engineering has been revolutionized by the evolution of a bacterial adaptive immune system, CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) into a radical technology that is making an expeditious progress in its mechanism, function and applicability.. METHODS A systematic literature review study was carried out with the help of all available information and online resources.. RESULTS In this review, we intend to elucidate different aspects of CRISPR in the light of current advancements. Utilizing a nonspecific Cas9 nuclease and a sequence specific programmable CRISPR RNA (crRNA), this system cleaves the target DNA with high precision. With a vast potential for profound implications, CRISPR has emerged as a mainstream method for plausible genomic manipulations in a range of organisms owing to its simplicity, accuracy and speed. A modified form of CRISPR system, known as CRISPR/Cpf1 that employs a smaller and simpler endonuclease...