Abstract
Currently, a new gene editing tool—the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) associated (Cas) system—is becoming a promising approach for genetic manipulation at the genomic level. This simple method, originating from the adaptive immune defense system in prokaryotes, has been developed and applied to antiviral research in humans. Based on the characteristics of virus-host interactions and the basic rules of nucleic acid cleavage or gene activation of the CRISPR-Cas system, it can be used to target both the virus genome and host factors to clear viral reservoirs and prohibit virus infection or replication. Here, we summarize recent progress of the CRISPR-Cas technology in editing host genes as an antiviral strategy.
Highlights
The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas immune system involves the helicase—called the Cas protein—which can bind to RNA transcribed from palindromic repeats of DNA or cleaved DNA paired with RNA spacers—which are transcripts from the short stretches of DNA
Thereafter, Zhang’s group described a structure guided optimization of the CRISPR-Cas9 complex to induce efficient transcriptional activation of up to ten endogenous genes at one time. This system contains a previously reported dCas9-VP64 fusion protein and an engineered single guide RNA (sgRNA) containing substitutions in the tetraloop and stem loop 2 domains of two minimal hairpin aptamers that selectively interacts with dimerized MS2 bacteriophage coat protein, the MS2 fused with the NF-κB trans-activating subunit p65 and the activation domain from human heat-shock factor 1 (HSF1)
CRISPR-Cas based targeting of host genes represents an alternative solution for virus related disease treatment for future applications
Summary
The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) system was first discovered as an immune response against bacteriophage infections and invading plasmids in prokaryotes such as Bacteria and Archaea [1,2]. Thereafter, Zhang’s group described a structure guided optimization of the CRISPR-Cas complex to induce efficient transcriptional activation of up to ten endogenous genes at one time This system contains a previously reported dCas9-VP64 fusion protein and an engineered sgRNA containing substitutions in the tetraloop and stem loop 2 domains of two minimal hairpin aptamers that selectively interacts with dimerized MS2 bacteriophage (a bacterial virus) coat protein, the MS2 fused with the NF-κB trans-activating subunit p65 and the activation domain from human heat-shock factor 1 (HSF1). Zhang’s group first experimentally confirmed its application as a genome editing tool in mammalian cells Distinct from both SpCas and SaCas, Cpf is a single RNA-guided endonuclease without tracrRNA and recognizes PAMs of.
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