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Delivery methods of CRISPR/Cas-mediated genome editing system
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Zahra Kordzadeh-Kermani    , Reza Kheirandish * , Hosseinali Sasan |
| Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran , kheirandish@uk.ac.ir |
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Abstract: (891 Views) |
CRISPR systems or clustered regularly interspaced short palindromic repeats and related proteins are natural microbial immune systems, which have been used as a tool for gene editing. To produce therapeutic effects, CRISPR components must be delivered directly to the target cells. There are different methods of delivery. In this study, some delivery systems through the cell membrane have been reviewed, including viral and non-viral delivery systems. The viral delivery system is the use of viruses such as adeno-associated virus vectors but there are concerns in its clinical use. The non-viral delivery system consists of various methods, including physical delivery and chemical delivery. In the physical delivery; the dose, duration, and specificity of the transfer are controlled more precisely, which includes the methods of electroporation, hydrodynamic injection, and microinjection. In the electroporation method, irreversible changes may occur in the physiology of the membrane, which threatens the cell's viability. In hydrodynamic injection, the volume required to start the injection is large and it is not a suitable method for human applications. Microinjection has high efficiency and precisely controlled dose, but it is very impractical for high-capacity cases. Although physical approaches are typically very successful in the laboratory, they generally do not scale well and are therefore less applicable for delivery to live conditions or high-throughput applications in the laboratory. In chemical delivery methods, instead of inducing a change in the target cell, the transferable cargo itself can be changed. There are two main forms of chemical delivery, including the method of encapsulation of the transferable substance and the method of modification of the transferable substance. Encapsulation can protect the cargo from enzymatic degradation or immune responses and enhance the specificity of delivery. Modification does not protect against degradation by proteases, nor are they capable of specific cell targeting. Therefore, successful use needs to be combined with encapsulation or other delivery methods. Considering that each delivery method has advantages and disadvantages, it is necessary to choose the most effective delivery method so that the CRISPR system be efficient for gene editing. |
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| Keywords: Cas9, CRISPR/Cas, Genome editing, Nonviral delivery, Viral delivery |
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Full-Text [PDF 799 kb]
(110 Downloads)
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Type of Study: Review |
Subject:
Animal
Received: 2023/02/11 | Accepted: 2023/05/26 | Published: 2023/08/29
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