RNA-directed gene editing specifically eradicates latent and prevents new HIV-1 infection

Hu, Wenhui; Kaminski, Rafal; Yang, Fan; Zhang, Yonggang; Cosentino, Laura; Li, Fang; Luo, Biao; Alvarez-Carbonell, David; García-Mesa, Yoelvis; Karn, Jonathan; Mo, Xianming; Khalili, Kamel

doi:10.1073/pnas.1405186111

Cited by 486 publications

(515 citation statements)

References 42 publications

(45 reference statements)

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“…In eukaryotes, the DSBs are more commonly repaired by the mechanism of error-prone non-homologous end joining (NHEJ), therefore generating sequence changes, for instance insertions and deletions (indels), around the DSBs . Owing to the simplicity of manipulation and versatility, the CRISPR/Cas9 system has been utilized as an attractive tool for various applications, such as genome-wide screening (Shalem et al, 2014;Zhou et al, 2014), gene repression and activation (Cheng et al, 2013;Doench et al, 2014;Gilbert et al, 2014), targeted fluorescence imaging (Tanenbaum et al, 2014) and novel approaches against pathogens including hepatitis B virus (Lin et al, 2014a;Seeger & Sohn, 2014), human papillomavirus (Kennedy et al, 2014), Epstein-Barr virus (Wang & Quake, 2014;Yuen et al, 2015), malaria (Wagner et al, 2014) and HIV-1 (Ebina et al, 2013;Hu et al, 2014;Ye et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

“…Using a pair of sgRNAs targeting the LTR of HIV-1, it was shown that HIV-1 provirus can be removed from the genome of infected cell lines (Ebina et al, 2013;Hu et al, 2014). By combining TALEN or CRISPR/Cas9 with PiggyBac technology, researches have generated inducedpluripotent stem cells (iPSC) homozygous for the naturally occurring CCR5 D32 variant resistant to HIV-1 infection .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9

Guan

et al. 2015

Journal of General Virology

189

139

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CCR5 serves as an essential coreceptor for human immunodeficiency virus type 1 (HIV-1) entry, and individuals with a CCR5 D32 variant appear to be healthy, making CCR5 an attractive target for control of HIV-1 infection. The CRISPR/Cas9, which functions as a naturally existing adaptive immune system in prokaryotes, has been recently harnessed as a novel nuclease system for genome editing in mammalian cells. Although CRISPR/Cas9 can be readily delivered into cell lines, due to the large size of the Cas9 protein, efficient delivery of CCR5-targeting CRISPR/Cas9 components into primary cells, including CD4 + T-cells, the primary target for HIV-1 infection in vivo, remains a challenge. In the current study, following design of a panel of top-ranked single-guided RNAs (sgRNAs) targeting the ORF of CCR5, we demonstrate that CRISPR/Cas9 can efficiently mediate the editing of the CCR5 locus in cell lines, resulting in the knockout of CCR5 expression on the cell surface. Next-generation sequencing revealed that various mutations were introduced around the predicted cleavage site of CCR5. For each of the three most effective sgRNAs that we analysed, no significant off-target effects were detected at the 15 top-scoring potential sites. More importantly, by constructing chimeric Ad5F35 adenoviruses carrying CRISPR/Cas9 components, we efficiently transduced primary CD4 + Tlymphocytes and disrupted CCR5 expression, and the positively transduced cells were conferred with HIV-1 resistance. To our knowledge, this is the first study establishing HIV-1 resistance in primary CD4 + T-cells utilizing adenovirus-delivered CRISPR/Cas9.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9

Guan

et al. 2015

Journal of General Virology

189

139

View full text Add to dashboard Cite

show abstract

“…Recently, several research groups have successfully applied the type II CRISPR systemSpCas9 protein from Streptococcus pyogenes with guided RNA (gRNA)-for targeted genome editing in diverse cell types and organisms, including human cells [12][13][14] . Most recently, a couple of studies have demonstrated the excision of the HIV-1 provirus from the host cell genome using gene-editing tools [15][16][17][18] . Here we apply the CRISPR/Cas9 system to directly target and disrupt the reverse-transcribed products of the lentiviral RNA genome during their life cycle within host cells.…”

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confidence: 99%

Use of the CRISPR/Cas9 system as an intracellular defense against HIV-1 infection in human cells

et al. 2015

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“…This proviral targeting strategy was mimicked with both TALENs [132] against integrated lentiviral LTRs and CRISPR/Cas9 in single and multiple configuration [133]. The authors identified targets within the viral U3 LTR that resulted in excision of the entire 9709 bp proviral HIV DNA and prevented reinfection within latently infected T cells [133]. …”

Section: Applicationsmentioning

confidence: 99%

Genome-Edited T Cell Therapies

Delhove

Qasim

2017

Curr Stem Cell Rep

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Purpose of ReviewAlternative approaches to conventional drug-based cancer treatments have seen T cell therapies deployed more widely over the last decade. This is largely due to their ability to target and kill specific cell types based on receptor recognition. Introduction of recombinant T cell receptors (TCRs) using viral vectors and HLA-independent T cell therapies using chimeric antigen receptors (CARs) are discussed. This article reviews the tools used for genome editing, with particular emphasis on the applications of site-specific DNA nuclease mediated editing for T cell therapies.Recent FindingsGenetic engineering of T cells using TCRs and CARs with redirected antigen-targeting specificity has resulted in clinical success of several immunotherapies. In conjunction, the application of genome editing technologies has resulted in the generation of HLA-independent universal T cells for allogeneic transplantation, improved T cell sustainability through knockout of the checkpoint inhibitor, programmed cell death protein-1 (PD-1), and has shown efficacy as an antiviral therapy through direct targeting of viral genomic sequences and entry receptors.SummaryThe combined use of engineered antigen-targeting moieties and innovative genome editing technologies have recently shown success in a small number of clinical trials targeting HIV and hematological malignancies and are now being incorporated into existing strategies for other immunotherapies.

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RNA-directed gene editing specifically eradicates latent and prevents new HIV-1 infection

Cited by 486 publications

References 42 publications

Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9

Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9

Use of the CRISPR/Cas9 system as an intracellular defense against HIV-1 infection in human cells

Genome-Edited T Cell Therapies

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