Efficient Inhibition of HIV Using CRISPR/Cas13d Nuclease System

Nguyen, Hoang; Wilson, Henrietta; Jayakumar, Sahana; Kulkarni, Viraj; Kulkarni, S. R.

doi:10.3390/v13091850

Cited by 18 publications

(13 citation statements)

References 77 publications

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“…Further, they reported that RfxCas13d (CasRx) significantly and efficiently inhibited the HIV-1 replication when targeting four separate, non-overlapping sites in the HIV-1 transcript in all the cell line models as well in primary CD4 + T-cells. Therefore their study concluded that the use of the CRISPR/Cas13d nuclease system to target acute and latent HIV infection will endow with an unconventional treatment modality against HIV ( Nguyen et al, 2021 ).…”

Section: Various Applications Of Cas13d In Basic Research and Biomedi...mentioning

confidence: 99%

Cas13d: A New Molecular Scissor for Transcriptome Engineering

Gupta¹,

Ghosh²,

Chakravarti³

et al. 2022

Front. Cell Dev. Biol.

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The discovery of Clustered Regularly Interspaced Palindromic Repeats (CRISPR) and its associated Cas endonucleases in bacterial and archaeal species allowed scientists to modify, utilized, and revolutionize this tool for genetic alterations in any species. Especially the type II CRISPR-Cas9 system has been extensively studied and utilized for precise and efficient DNA manipulation in plant and mammalian systems over the past few decades. Further, the discovery of the type V CRISPR-Cas12 (Cpf1) system provides more flexibility and precision in DNA manipulation in prokaryotes, plants, and animals. However, much effort has been made to employ and utilize the above CRISPR tools for RNA manipulation but the ability of Cas9 and Cas12 to cut DNA involves the nuisance of off-target effects on genes and thus may not be employed in all RNA-targeting applications. Therefore, the search for new and diverse Cas effectors which can precisely detect and manipulate the targeted RNA begins and this led to the discovery of a novel RNA targeting class 2, type VI CRISPR-Cas13 system. The CRISPR-Cas13 system consists of single RNA-guided Cas13 effector nucleases that solely target single-stranded RNA (ssRNA) in a programmable way without altering the DNA. The Cas13 effectors family comprises four subtypes (a-d) and each subtype has distinctive primary sequence divergence except the two consensuses Higher eukaryotes and prokaryotes nucleotide-binding domain (HEPN) that includes RNase motifs i.e. R-X4-6-H. These two HEPN domains are solely responsible for executing targetable RNA cleavage activity with high efficiency. Further, recent studies have shown that Cas13d exhibits higher efficiency and specificity in cleaving targeted RNA in the mammalian system compared to other Cas13 endonucleases of the Cas13 enzyme family. In addition to that, Cas13d has shown additional advantages over other Cas13 variants, structurally as well as functionally which makes it a prominent and superlative tool for RNA engineering and editing. Therefore considering the advantages of Cas13d over previously characterized Cas13 subtypes, in this review, we encompass the structural and mechanistic properties of type VI CRISPR-Cas13d systems, an overview of the current reported various applications of Cas13d, and the prospects to improve Cas13d based tools for diagnostic and therapeutic purposes.

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Section: Various Applications Of Cas13d In Basic Research and Biomedi...mentioning

confidence: 99%

Cas13d: A New Molecular Scissor for Transcriptome Engineering

Gupta¹,

Ghosh²,

Chakravarti³

et al. 2022

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

show abstract

“…While Cas9 produces blunt double-strand cuts, Cas12a produces staggered cuts in dsDNA. Additionally, the Cas13 nuclease which targets the RNA has been used in HIV-1 infected cells with significant results [ 31 ]. With the discovery of multiple nuclease variants, a varied usage of these Cas proteins can be employed in gene editing systems for getting effective results.…”

Section: Crispr/cas System In Genome Editingmentioning

confidence: 99%

“…The RNA-editing Cas13 system has been recently tested against HIV-1 infected cells. The Cas13d system was able to effectively inhibit HIV-1 infection in primary CD4 + T cells and also, suppressed reactivated HIV-1 from latently infected cells [ 31 ]. The CRISPR-Cas13a targets HIV-1 RNA leading to a reduction in viral gene expression.…”

Section: Crispr/cas Mediated Inhibition Of Hiv-1mentioning

confidence: 99%

CRISPR/Cas9: a tool to eradicate HIV-1

Bhowmik

Chaubey

2022

AIDS Res Ther

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The development of antiretroviral therapy (ART) has been effective in suppressing HIV replication. However, severe drug toxicities due to the therapy and its failure in targeting the integrated proviral genome have led to the introduction of a new paradigm of gene-based therapies. With its effective inhibition and high precision, clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease (Cas9) or CRISPR/Cas9 has emerged as an effective genome editing tool in the last decade. Mediated by guide RNAs (gRNAs), Cas9 endonuclease acts like genetic scissors that can modify specific target sites. With this concept, CRISPR/Cas9 has been used to target the integrated proviral HIV-1 genome both in in vitro as well as in vivo studies including non-human primates. The CRISPR has also been tested for targeting latent HIV-1 by modulating the proviral transcription with the help of a specialized Cas9 mutant. Overcoming the limitations of the current therapy, CRISPR has the potential to become the primary genome editing tool for eradicating HIV-1 infection. In this review, we summarize the recent advancements of CRISPR to target the proviral HIV-1 genome, the challenges and future prospects.

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“…Because Cas12a and, especially, its orthologs Cas12j and Cas12f are much smaller than Cas9, their improved delivery with size-restricted vehicles, such as AAV, viral, and nonviral particles ( Briolay et al., 2021 ), is highly probable and should motivate their development and application in the future. As alternatives to the genetic and epigenetic silencing of HIV, two recent reports demonstrated the efficient suppression of HIV replication and reactivation at the mRNA/gRNA level with Cas13a ( Yin et al., 2020 ) and Cas13d ( Nguyen et al., 2021 ). Interestingly, Cas13a degraded gRNA packed into viral particles.…”

Section: Perspectives In Crispr/cas Anti-hiv Therapy and Concluding R...mentioning

confidence: 99%

Application of CRISPR/Cas Genomic Editing Tools for HIV Therapy: Toward Precise Modifications and Multilevel Protection

Maslennikova

Mazurov

2022

Front. Cell. Infect. Microbiol.

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Although highly active antiretroviral therapy (HAART) can robustly control human immunodeficiency virus (HIV) infection, the existence of latent HIV in a form of proviral DNA integrated into the host genome makes the virus insensitive to HAART. This requires patients to adhere to HAART for a lifetime, often leading to drug toxicity or viral resistance to therapy. Current genome-editing technologies offer different strategies to reduce the latent HIV reservoir in the body. In this review, we systematize the research on CRISPR/Cas-based anti-HIV therapeutic methods, discuss problems related to viral escape and gene editing, and try to focus on the technologies that effectively and precisely introduce genetic modifications and confer strong resistance to HIV infection. Particularly, knock-in (KI) approaches, such as mature B cells engineered to produce broadly neutralizing antibodies, T cells expressing fusion inhibitory peptides in the context of inactivated viral coreceptors, or provirus excision using base editors, look very promising. Current and future advancements in the precision of CRISPR/Cas editing and its delivery will help extend its applicability to clinical HIV therapy.

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Efficient Inhibition of HIV Using CRISPR/Cas13d Nuclease System

Cited by 18 publications

References 77 publications

Cas13d: A New Molecular Scissor for Transcriptome Engineering

Cas13d: A New Molecular Scissor for Transcriptome Engineering

CRISPR/Cas9: a tool to eradicate HIV-1

Application of CRISPR/Cas Genomic Editing Tools for HIV Therapy: Toward Precise Modifications and Multilevel Protection

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