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Non-VLPs
Published in Paul Pumpens, Single-Stranded RNA Phages, 2020
Concerning the immune activity of the CRISPR system against RNA phages, a breakthrough investigation was performed by Virginijus Šikšnys’ excellent team (Tamulaitis et al. 2014). By their investigation of nucleic acid specificity and mechanism of CRISPR interference for the Streptococcus thermophilus complex, the authors found that the type III-A StCsm complex targeted RNA and not DNA. When expressed in E. coli, the StCsm complex restricted the phage MS2 in a Csm3 nuclease-dependent manner. It was stressed by the authors that the phage MS2 is a preferable model to investigate RNA targeting by the CRISPR-Cas system in vivo because no DNA intermediate is formed during the life cycle of this phage. The pCRISPR_MS2 plasmid carried the synthetic CRISPR array of five repeats interspaced by four 36-nucleotide spacers targeting correspondingly the maturation, lysis, coat, and replicase sequences of MS2 RNA. The data demonstrated clearly that the StCsm complex conveyed in vivo resistance to the RNA phage MS2 in the heterologous E. coli host (Tamulaitis et al. 2014).
TYK2 as a novel therapeutic target in psoriasis
Published in Expert Review of Clinical Pharmacology, 2023
Sarah Elyoussfi, Shraddha S Rane, Steve Eyre, Richard B Warren
Experimental techniques include investigation of how transcription factors and other proteins interact with DNA (ChIP), chromatin structure and interactions (3C), and gene expression (eQTL and reporter gene assays) in the relevant cell types. Methods such as Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein 9 (CRISPR/Cas9) genome-editing systems can evaluate the effect of altering individual SNP alleles on gene expression [39]. By using relevant cell types, such as T-cells or keratinocytes, the effect of regulatory genome regions on subsequent gene expression can be examined using reporter gene assays. CRISPR interference systems can be used to downregulate or upregulate transcription of these regulatory regions. Informative functional readouts are then used to help us understand the function of disease-associated variants and the biological pathways on which they act [40].
An overview: CRISPR/Cas-based gene editing for viral vaccine development
Published in Expert Review of Vaccines, 2022
Santosh Bhujbal, Rushikesh Bhujbal, Prabhanjan Giram
Multiple viral infections cause a latent infection in which the viral gene is retained in a dormant state with periodic reactivation due to overexpression suppression. Because, dormancy is among the characteristics of HIV type 1 is particularly crucial, as dormancy is one of the primary hurdles to HIV treatment. The reactivation of HIV-1 out of its dormant condition, making individuals receptive to antiretroviral treatment, might be a successful therapeutic technique. For specific CRISPR activation (CRISPRa) and CRISPR interference (CRISPRi), researchers employed modified Cas9 lacking nuclease activity combined with transcriptional activators and repressors, respectively. Cas9 modifications with alterations in its two endonuclease regions inhibit Cas9’s nuclease function, allowing the development of programmed RNA-dependent DNA-binding enzymes [122]. dCas9 has been effectively coupled with various protein or active regions to generate a specific expression of genes or cellular targeting strategies [123].
Common therapeutic advances for Duchenne muscular dystrophy (DMD)
Published in International Journal of Neuroscience, 2021
Arash Salmaninejad, Yousef Jafari Abarghan, Saeed Bozorg Qomi, Hadi Bayat, Meysam Yousefi, Sara Azhdari, Samaneh Talebi, Majid Mojarrad
According to, autologous iPS cells can be a potential therapeutic strategy for efficiently and safely regeneration of muscles in patients with DMD without causing immune activation. CRISPR-Cas9 system has recently modified and repurposed for genome regulation instead of genome editing [184]. To do this, endonuclease domains of the Cas9 protein have been inactivated and created a programmable RNA-dependent DNA-binding proteins called dead Cas9 or (dCas9). Dead Cas9 proteins cannot cleave DNA but enabling CRISPR-Cas9 system to transcriptionally activate or suppress of gene expression and also allowing regulation of endogenous epigenome via fusing a repressive or activating effector to dCas9 protein. According to, this combined protein will be directed to a specific site of the genome and therefore able to conduction of repression or activation of a specific gene expression [184]. This modified type of CRISPR system named as CRISPR interference (CRISPRi) [184].