Modeling human disease in rodents by CRISPR/Cas9 genome editing (original) (raw)
Related papers
Creating cell and animal models of human disease by genome editing using CRISPR/Cas9
The Journal of Gene Medicine, 2019
Aims: Set of unique sequences in bacterial genomes, responsible for protecting bacteria against bacteriophages, have been recently used for genetic manipulation of specific points in the genome. These system comprises of one RNA component and one enzyme component, known as CRISPR and Cas9, respectively. The present review focuses on the applications of the CRISPR/Cas9 technology in the development of cellular and animal models of human disease. Discussion: Making a desired genetic alteration depend on the design of RNA molecules that guide endonucleases to a favorable genomic location. With the discovery of the CRISPR/Cas9 technology, scientists are able to achieve higher levels of accuracy as it possesses advantages over the alternative methods for editing genome including simple design, high targeting efficiency and the ability to create simultaneous alterations in multiple sequences. These peruse the researchers to apply this technology for creating cellular and animal models of human diseases by knock-in, knockout and Indel mutation strategies such as Huntington, Cardiovascular disorders, and cancers. Conclusion: Optimized CRISPR/Cas9 technology will facilitate access to valuable novel cellular and animal genetic models with applications in development of innovative drug discovery and gene therapy.
European chemical bulletin , 2023
Through the exact change of live creatures' genetic makeup, genome editing technologies have revolutionised the area of biomedicine. CRISPR-Cas9 has distinguished itself among these technologies as a potent tool with enormous potential for modelling diseases and discovering new drugs. The use of CRISPR-Cas9 in biomedical research is examined in this work, with a particular emphasis on how it might be used to produce new drugs and model diseases. We go over the CRISPR-Cas9 tenets, why it's better than other genome editing techniques, and how it's been used successfully to treat various diseases in various disease models. We also emphasise how CRISPR-Cas9 has accelerated the identification of fresh therapeutic targets and the creation of more potent medications. We also talk about the problems and moral issues that CRISPR-Cas9 technology raises, putting a focus on its proper and moral application. Overall, this study highlights the important contributions of CRISPR-Cas9 to disease modelling and medication discovery, highlighting its potential to fundamentally alter the area of biomedicine.
CRISPR/Cas9-mediated Genome Editing: In vivo Review
Journal of Lab Animal Research
The CRISPR/Cas9 system has been a game-changer in genetics and biotechnology. This study aimed to investigate the existing in vivo uses and their potential to increase our understanding of gene function and biological processes in animal models. With its remarkable precision and accuracy, researchers can now easily edit specific genes within cells and organisms. This technology has opened up new avenues for studying genetic diseases and developing therapies to treat them. One of the most significant advantages of the CRISPR/Cas9 system is its ability to create precise cellular and animal models of human diseases. This allows researchers to investigate the role of genetics in disease development and to develop more effective therapies. For example, the system can correct genetic mutations that cause cystic fibrosis or sickle cell anemia. The therapeutic potential of CRISPR/Cas9 is enormous, especially in gene therapy. By correcting specific genetic mutations, the system can potential...
Applying the CRISPR/Cas9 for treating human and animal diseases: a comprehensive review
Annals of Animal Science
Recently, genome editing tools have been extensively used in many biomedical sciences. The gene editing system is applied to modify the DNA sequences in the cellular system to comprehend their physiological response. A developing genome editing technology like clustered regularly short palindromic repeats (CRISPR) is widely expended in medical sciences. CRISPR and CRISPR-associated protein 9 (CRISPR/Cas9) system is being exploited to edit any DNA mutations related to inherited ailments to investigate in animals (in vivo) and cell lines (in vitro). Remarkably, CRISPR/Cas9 could be employed to examine treatments of many human genetic diseases such as Cystic fibrosis, Tyrosinemia, Phenylketonuria, Muscular dystrophy, Parkinson’s disease, Retinoschisis, Hemophilia, β-Thalassemia and Atherosclerosis. Moreover, CRISPR/Cas9 was used for disease resistance such as Tuberculosis, Johne’s diseases, chronic enteritis, and Brucellosis in animals. Finally, this review discusses existing progress ...
Rapid generation of mouse models with defined point mutations by the CRISPR/Cas9 system
Scientific reports, 2014
Introducing a point mutation is a fundamental method used to demonstrate the roles of particular nucleotides or amino acids in the genetic elements or proteins, and is widely used in in vitro experiments based on cultured cells and exogenously provided DNA. However, the in vivo application of this approach by modifying genomic loci is uncommon, partly due to its technical and temporal demands. This leaves many in vitro findings un-validated under in vivo conditions. We herein applied the CRISPR/Cas9 system to generate mice with point mutations in their genomes, which led to single amino acid substitutions in proteins of interest. By microinjecting gRNA, hCas9 mRNA and single-stranded donor oligonucleotides (ssODN) into mouse zygotes, we introduced defined genomic modifications in their genome with a low cost and in a short time. Both single gRNA/WT hCas9 and double nicking set-ups were effective. We also found that the distance between the modification site and gRNA target site was ...
CRISPR-Cas9 knockin mice for genome editing and cancer modeling
Cell, 2014
CRISPR-Cas9 is a versatile genome editing technology for studying the functions of genetic elements. To broadly enable the application of Cas9 in vivo, we established a Cre-dependent Cas9 knockin mouse. We demonstrated in vivo as well as ex vivo genome editing using adeno-associated virus (AAV)-, lentivirus-, or particle-mediated delivery of guide RNA in neurons, immune cells, and endothelial cells. Using these mice, we simultaneously modeled the dynamics of KRAS, p53, and LKB1, the top three significantly mutated genes in lung adenocarcinoma. Delivery of a single AAV vector in the lung generated loss-of-function mutations in p53 and Lkb1, as well as homology-directed repair-mediated Kras(G12D) mutations, leading to macroscopic tumors of adenocarcinoma pathology. Together, these results suggest that Cas9 mice empower a wide range of biological and disease modeling applications.
CRISPR-Cas9 gene editing and human diseases
bioinformation, 2022
The author's state that they adhere with COPE guidelines on publishing ethics as described elsewhere at https://publicationethics.org/. The authors also undertake that they are not associated with any other third party (governmental or non-governmental agencies) linking with any form of unethical issues connecting to this publication. The authors also declare that they are not withholding any information that is misleading to the publisher in regard to this article.
CRISPR/Cas based gene editing: marking a new era in medical science
Molecular Biology Reports, 2021
CRISPR/Cas9 system, a bacterial adaptive immune system developed into a genome editing technology, has emerged as a powerful tool revolutionising genome engineering in all branches of biological science including agriculture, research and medicine. Rapid evolution of CRISPR/Cas9 system from the generation of double strand breaks to more advanced applications on gene regulation has made the wide-spread use of this technology possible. Medical science has benefited greatly from CRISPR/Cas9; being both a versatile and economical tool, it has brought gene therapy closer to reality. In this review, the development of CRISPR/Cas9 system, variants thereof and its application in different walks of medical science- research, diagnostics and therapy, will be discussed.
Genome editing: A perspective on the application of CRISPR/Cas9 to study human diseases (Review)
International Journal of Molecular Medicine, 2019
Genome editing reemerged in 2012 with the development of cRISPR/cas9 technology, which is a genetic manipulation tool derived from the defense system of certain bacteria against viruses and plasmids. This method is easy to apply and has been used in a wide variety of experimental models, including cell lines, laboratory animals, plants, and even in human clinical trials. The cRISPR/cas9 system consists of directing the cas9 nuclease to create a site-directed double-strand dNA break using a small RNA molecule as a guide. A process that allows a permanent modification of the genomic target sequence can repair the damage caused to dNA. In the present study, the basic principles of the cRISPR/cas9 system are reviewed, as well as the strategies and modifications of the enzyme cas9 to eliminate the off-target cuts, and the different applications of cRISPR/cas9 as a system for visualization and gene expression activation or suppression. In addition, the review emphasizes on the potential application of this system in the treatment of different diseases, such as pulmonary, gastrointestinal, hematologic, immune system, viral, autoimmune and inflammatory diseases, and cancer.
A Mouse Geneticist’s Practical Guide to CRISPR Applications
Genetics, 2014
CRISPR/Cas9 system of RNA-guided genome editing is revolutionizing genetics research in a wide spectrum of organisms. Even for the laboratory mouse, a model that has thrived under the benefits of embryonic stem (ES) cell knockout capabilities for nearly three decades, CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 technology enables one to manipulate the genome with unprecedented simplicity and speed. It allows generation of null, conditional, precisely mutated, reporter, or tagged alleles in mice. Moreover, it holds promise for other applications beyond genome editing. The crux of this system is the efficient and targeted introduction of DNA breaks that are repaired by any of several pathways in a predictable but not entirely controllable manner. Thus, further optimizations and improvements are being developed. Here, we summarize current applications and provide a practical guide to use the CRISPR/Cas9 system for mouse mutagenesis, based on published report...