A combinational CRISPR/Cas9 gene-editing approach can halt HIV replication and prevent viral escape DOI Creative Commons
Robert Jan Lebbink,

Dorien C. M. de Jong,

Femke Wolters

et al.

Scientific Reports, Journal Year: 2017, Volume and Issue: 7(1)

Published: Feb. 8, 2017

Abstract HIV presents one of the highest evolutionary rates ever detected and combination antiretroviral therapy is needed to overcome plasticity virus population control viral replication. Conventional treatments lack ability clear latent reservoir, which remains major obstacle towards a cure. Novel strategies, such as CRISPR/Cas9 gRNA-based genome-editing, can permanently disrupt genome. However, genome-editing may accelerate escape, questioning feasibility approach. Here, we demonstrate that targeting single loci, only partially inhibits replication facilitates rapid escape at target site. A combinatorial approach two strong gRNAs different regions genome completely abrogate prevent escape. Our data shows accelerating effect gene-editing on be provide future alternative for HIV-infection.

Language: Английский

Applications of CRISPR technologies in research and beyond DOI
Rodolphe Barrangou, Jennifer A. Doudna

Nature Biotechnology, Journal Year: 2016, Volume and Issue: 34(9), P. 933 - 941

Published: Sept. 1, 2016

Language: Английский

Citations

870
A century of the phage: past, present and future DOI
George P. C. Salmond, Peter C. Fineran

Nature Reviews Microbiology, Journal Year: 2015, Volume and Issue: 13(12), P. 777 - 786

Published: Nov. 9, 2015

Language: Английский

Citations

666
Endosomal Escape and Delivery of CRISPR/Cas9 Genome Editing Machinery Enabled by Nanoscale Zeolitic Imidazolate Framework DOI
Shahad K. Alsaiari, Sachin Patil, Mram Alyami

et al.

Journal of the American Chemical Society, Journal Year: 2017, Volume and Issue: 140(1), P. 143 - 146

Published: Dec. 22, 2017

CRISPR/Cas9 is a combined protein (Cas9) and an engineered single guide RNA (sgRNA) genome editing platform that offers revolutionary solutions to genetic diseases. It has, however, double delivery problem owning the large size highly charged component. In this work, we report first example of encapsulated by nanoscale zeolitic imidazole frameworks (ZIFs) with loading efficiency 17% enhanced endosomal escape promoted protonated moieties. The gene potential ZIF-8 (CC-ZIFs) further verified knocking down expression green fluorescent 37% over 4 days. CC-ZIFs are biocompatible easily scaled-up offering excellent capacity controlled codelivery intact Cas9 sgRNA.

Language: Английский

Citations

454
In Vivo Target Gene Activation via CRISPR/Cas9-Mediated Trans-epigenetic Modulation DOI Creative Commons

Hsin‐Kai Liao,

Fumiyuki Hatanaka, Toshikazu Araoka

et al.

Cell, Journal Year: 2017, Volume and Issue: 171(7), P. 1495 - 1507.e15

Published: Dec. 1, 2017

Language: Английский

Citations

402
Latest Developed Strategies to Minimize the Off-Target Effects in CRISPR-Cas-Mediated Genome Editing DOI Creative Commons
Muhammad Naeem,

Saman Majeed,

Mubasher Zahir Hoque

et al.

Cells, Journal Year: 2020, Volume and Issue: 9(7), P. 1608 - 1608

Published: July 2, 2020

Gene editing that makes target gene modification in the genome by deletion or addition has revolutionized era of biomedicine. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 emerged as a substantial tool due to its simplicity use, less cost and extraordinary efficiency than conventional gene-editing tools, including zinc finger nucleases (ZFNs) Transcription activator-like effector (TALENs). However, potential off-target activities are crucial shortcomings CRISPR system. Numerous types approaches have been developed reduce effects. Here, we review several latest effects, biased unbiased detection, cytosine adenine base editors, prime editing, dCas9, Cas9 paired nickase, ribonucleoprotein (RNP) delivery truncated gRNAs. This article provides extensive information cautiously interpret effects assist basic clinical applications

Language: Английский

Citations

367
Engineering the Delivery System for CRISPR-Based Genome Editing DOI Creative Commons
Zachary Glass, Matthew Lee, Yamin Li

et al.

Trends in biotechnology, Journal Year: 2018, Volume and Issue: 36(2), P. 173 - 185

Published: Jan. 2, 2018

Language: Английский

Citations

305
Elimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing DOI Creative Commons
Rafal Kaminski, Yilan Chen, Tracy Fischer

et al.

Scientific Reports, Journal Year: 2016, Volume and Issue: 6(1)

Published: March 4, 2016

Abstract We employed an RNA-guided CRISPR/Cas9 DNA editing system to precisely remove the entire HIV-1 genome spanning between 5′ and 3′ LTRs of integrated proviral copies from latently infected human CD4+ T-cells. Comprehensive assessment whole-genome sequencing eradicated cells ruled out any off-target effects by our technology that might compromise integrity host further showed no effect on several cell health indices including viability, cycle apoptosis. Persistent co-expression Cas9 specific targeting guide RNAs in HIV-1-eradicated T-cells protected them against new infection HIV-1. Lentivirus-delivered significantly diminished replication primary T-cell cultures drastically reduced viral load ex vivo culture obtained patients. Thus, gene using may provide a therapeutic path for eliminating potentially serve as novel effective platform toward curing AIDS.

Language: Английский

Citations

301
CRISPR/Cas9 editing of endogenous banana streak virus in the B genome of Musa spp. overcomes a major challenge in banana breeding DOI Creative Commons
Jaindra Nath Tripathi, Valentine Otang Ntui,

Mily Ron

et al.

Communications Biology, Journal Year: 2019, Volume and Issue: 2(1)

Published: Jan. 31, 2019

Abstract Presence of the integrated endogenous banana streak virus (eBSV) in B genome plantain (AAB) is a major challenge for breeding and dissemination hybrids. As eBSV activates into infectious viral particles under stress, progenitor Musa balbisiana its derivants, having at least one genome, cannot be used as parents crop improvement. Here, we report strategy to inactivate by editing sequences. The regenerated genome-edited events Gonja Manjaya showed mutations targeted sites with potential prevent proper transcription or/and translational functional proteins. Seventy-five percent edited remained asymptomatic comparison non-edited control plants water stress conditions, confirming inactivation particles. This study paves way improvement germplasm use programs produce hybrids that can globally disseminated.

Language: Английский

Citations

278
In Vivo Excision of HIV-1 Provirus by saCas9 and Multiplex Single-Guide RNAs in Animal Models DOI Creative Commons
Chaoran Yin, Ting Zhang,

Xiying Qu

et al.

Molecular Therapy, Journal Year: 2017, Volume and Issue: 25(5), P. 1168 - 1186

Published: March 31, 2017

CRISPR-associated protein 9 (Cas9)-mediated genome editing provides a promising cure for HIV-1/AIDS; however, gene delivery efficiency in vivo remains an obstacle to overcome. Here, we demonstrate the feasibility and of excising HIV-1 provirus three different animal models using all-in-one adeno-associated virus (AAV) vector deliver multiplex single-guide RNAs (sgRNAs) plus Staphylococcus aureus Cas9 (saCas9). The quadruplex sgRNAs/saCas9 outperformed duplex integrated cultured neural stem/progenitor cells from Tg26 transgenic mice. Intravenously injected AAV-DJ/8 excised proviral DNA significantly reduced viral RNA expression several organs/tissues In EcoHIV acutely infected mice, intravenously systemic infection, as determined by live bioluminescence imaging. Additionally, this induced efficient excision, PCR genotyping liver, lungs, brain, spleen. Finally, humanized bone marrow/liver/thymus (BLT) mice with chronic successful excision was detected spleen, heart, colon, brain after single intravenous injection AAV-DJ/8. conclusion, solid tissues/organs can be achieved via AAV delivery, significant step toward human clinical trials.Graphical abstract

Language: Английский

Citations

272
CRISPR/Cas9-Mediated Genome Editing of Herpesviruses Limits Productive and Latent Infections DOI Creative Commons

Ferdy R. van Diemen,

Elisabeth Kruse,

Marjolein J. G. Hooykaas

et al.

PLoS Pathogens, Journal Year: 2016, Volume and Issue: 12(6), P. e1005701 - e1005701

Published: June 30, 2016

Herpesviruses infect the majority of human population and can cause significant morbidity mortality. Herpes simplex virus (HSV) type 1 causes cold sores herpes keratitis, whereas HSV-2 is responsible for genital herpes. Human cytomegalovirus (HCMV) most common viral congenital defects serious disease in immuno-compromised individuals. Epstein-Barr (EBV) associated with infectious mononucleosis a broad range malignancies, including Burkitt’s lymphoma, nasopharyngeal carcinoma, Hodgkin’s disease, post-transplant lymphomas. persist their host life by establishing latent infection that interrupted periodic reactivation events during which replication occurs. Current antiviral drug treatments target clinical manifestations this productive stage, but they are ineffective at eliminating these viruses from infected host. Here, we set out to combat both herpesvirus infections exploiting CRISPR/Cas9 system genetic elements important fitness. We show effective abrogation HCMV HSV-1 targeting gRNAs essential genes. Simultaneous multiple completely abolished production particles cells. Using same approach, EBV be almost cleared latently EBV-transformed tumor Our studies indicate effectively targeted genomes as potent prophylactic therapeutic anti-viral strategy may used impair clear infection.

Language: Английский

Citations

262