HIV-1 inhibition in cells with CXCR4 mutant genome created by CRISPR-Cas9 and piggyBac recombinant technologies

Shuai, Liu; Wang, Qiankun; Yu, Xijie; Li, Yilin; Guo, Yandan; Liu, Zhepeng; Sun, Fuyun; Hou, Wei; Li, Chunmei; Wu, Li; Guo, Deyin; Chen, Shuliang

doi:10.1038/s41598-018-26894-4

Cited by 34 publications

(36 citation statements)

References 43 publications

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“…It is used in non-viral vectors for transgenesis 17 , 18 , gene therapy 7 , 19 , insertional mutagenesis 20 , and genetic screens 21 – 23 . It has also found application in novel therapeutic strategies including CAR T-cell engineering 24 – 26 , CRISPR/Cas-mediated gene therapy 27 – 29 , and human induced pluripotent stem cells (iPSC) engineering 30 – 32 . Useful variants have been developed through random mutation including a hyperactive pB transposase called hyPBase 33 as well as one that can excise pB but cannot integrate it 34 .…”

Section: Introductionmentioning

confidence: 99%

Structural basis of seamless excision and specific targeting by piggyBac transposase

et al. 2020

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The piggyBac DNA transposon is used widely in genome engineering applications. Unlike other transposons, its excision site can be precisely repaired without leaving footprints and it integrates specifically at TTAA tetranucleotides. We present cryo-EM structures of piggyBac transpososomes: a synaptic complex with hairpin DNA intermediates and a strand transfer complex capturing the integration step. The results show that the excised TTAA hairpin intermediate and the TTAA target adopt essentially identical conformations, providing a mechanistic link connecting the two unique properties of piggyBac. The transposase forms an asymmetric dimer in which the two central domains synapse the ends while two C-terminal domains form a separate dimer that contacts only one transposon end. In the strand transfer structure, target DNA is severely bent and the TTAA target is unpaired. In-cell data suggest that asymmetry promotes synaptic complex formation, and modifying ends with additional transposase binding sites stimulates activity.

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Section: Introductionmentioning

confidence: 99%

Structural basis of seamless excision and specific targeting by piggyBac transposase

et al. 2020

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“…As CXCR4 and its ligand CXC12 (SDF-1) play pivotal roles in hematopoietic/progenitor cells development and thymic differentiation (Nagasawa et al, 1996; Dar et al, 2006), we should consider the safety and side effects of clinical application by targeting CXCR4 in HSPCs. However, CXCR4-deficient human T cells remain functional in a mouse model (Chung et al, 2010; Yuan et al, 2012), and we also used CRISPR-Cas9 and piggyBac recombinant technologies to create CXCR4 P191A mutant with HIV-1 infection inhibition function and without deficiency of CXCR4 function(Liu S. et al, 2018), it is possible for CRISPR-Cas9 editing of CXCR4 in human mature post-thymic CD4+ T cells for the purpose of HIV-1/AIDS therapy.…”

Section: Crispr/cas9 Technology Application In Hiv-1/aids Treatmentmentioning

confidence: 99%

Application of CRISPR/Cas9-Based Gene Editing in HIV-1/AIDS Therapy

Xiao

Guo

Chen

2019

Front. Cell. Infect. Microbiol.

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132

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Despite the fact that great efforts have been made in the prevention and therapy of HIV-1 infection, HIV-1/AIDS remains a major threat to global human health. Highly active antiretroviral therapy (HAART) can suppress virus replication, but it cannot eradicate latent viral reservoirs in HIV-1/AIDS patients. Recently, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9) system has been engineered as an effective gene-editing technology with the potential to treat HIV-1/AIDS. It can be used to target cellular co-factors or HIV-1 genome to reduce HIV-1 infection and clear the provirus, as well as to induce transcriptional activation of latent virus in latent viral reservoirs for elimination. This versatile gene editing technology has been successfully applied to HIV-1/AIDS prevention and reduction in human cells and animal models. Here, we update the rapid progress of CRISPR/Cas9-based HIV-1/AIDS therapy research in recent years and discuss the limitations and future perspectives of its application.

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“…Modified cells showed resistance to CCR5-utilizing virus and a selective advantage over wild-type cells (Wang et al, 2014). Several studies have also reported successful inhibition of CXCR4 expression and a resultant increase in HIV-1 resistance in vitro (Hou et al, 2015; Liu et al, 2017, 2018; Wang et al, 2017). Recently, a study reported the successful ablation of both the CCR5 and CXCR4 co-receptors in vitro .…”

Section: Crispr Targeting Ccr5mentioning

confidence: 99%

Gene Editing of HIV-1 Co-receptors to Prevent and/or Cure Virus Infection

et al. 2018

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Antiretroviral therapy has prolonged the lives of people living with human immunodeficiency virus type 1 (HIV-1), transforming the disease into one that can be controlled with lifelong therapy. The search for an HIV-1 vaccine has plagued researchers for more than three decades with little to no success from clinical trials. Due to these failures, scientists have turned to alternative methods to develop next generation therapeutics that could allow patients to live with HIV-1 without the need for daily medication. One method that has been proposed has involved the use of a number of powerful gene editing tools; Zinc Finger Nucleases (ZFN), Transcription Activator–like effector nucleases (TALENs), and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 to edit the co-receptors (CCR5 or CXCR4) required for HIV-1 to infect susceptible target cells efficiently. Initial safety studies in patients have shown that editing the CCR5 locus is safe. More in depth in vitro studies have shown that editing the CCR5 locus was able to inhibit infection from CCR5-utilizing virus, but CXCR4-utilizing virus was still able to infect cells. Additional research efforts were then aimed at editing the CXCR4 locus, but this came with other safety concerns. However, in vitro studies have since confirmed that CXCR4 can be edited without killing cells and can confer resistance to CXCR4-utilizing HIV-1. Utilizing these powerful new gene editing technologies in concert could confer cellular resistance to HIV-1. While the CD4, CCR5, CXCR4 axis for cell-free infection has been the most studied, there are a plethora of reports suggesting that the cell-to-cell transmission of HIV-1 is significantly more efficient. These reports also indicated that while broadly neutralizing antibodies are well suited with respect to blocking cell-free infection, cell-to-cell transmission remains refractile to this approach. In addition to stopping cell-free infection, gene editing of the HIV-1 co-receptors could block cell-to-cell transmission. This review aims to summarize what has been shown with regard to editing the co-receptors needed for HIV-1 entry and how they could impact the future of HIV-1 therapeutic and prevention strategies.

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HIV-1 inhibition in cells with CXCR4 mutant genome created by CRISPR-Cas9 and piggyBac recombinant technologies

Cited by 34 publications

References 43 publications

Structural basis of seamless excision and specific targeting by piggyBac transposase

Structural basis of seamless excision and specific targeting by piggyBac transposase

Application of CRISPR/Cas9-Based Gene Editing in HIV-1/AIDS Therapy

Gene Editing of HIV-1 Co-receptors to Prevent and/or Cure Virus Infection

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