Strategies for controlling CRISPR/Cas9 off-target effects and biological variations in mammalian genome editing experiments

Kimberland, Michelle L.; Hou, Wangfang; Alfonso-Pecchio, Adolfo; Wilson, Stephen; Rao, Yanhua; Zhang, Shu; Lu, Quinn

doi:10.1016/j.jbiotec.2018.08.007

Cited by 73 publications

(57 citation statements)

References 108 publications

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“…By titrating the concentration of the Cas9 RNPs to an optimal level and limiting the window of exposure to the RNPs, OTEs in the edited cells can be substantially reduced by 2.2 to 19 folds [91,93,95,96]. Similar to using Cas9 mRNA, cellular toxicity associated with exogenous DNA and the possibility of foreign DNA integration can be avoided with Cas9 RNP [97]. DNA repair pathways…”

Section: Modality Of the Crispr/cas9 System During Deliverymentioning

confidence: 99%

Mitigating off-target effects in CRISPR/Cas9-mediated in vivo gene editing

2020

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The rapid advancement of genome editing technologies has opened up new possibilities in the field of medicine. Nuclease-based techniques such as the CRISPR/Cas9 system are now used to target genetically linked disorders that were previously hard-totreat. The CRISPR/Cas9 gene editing approach wields several advantages over its contemporary editing systems, notably in the ease of component design, implementation and the option of multiplex genome editing. While results from the early phase clinical trials have been encouraging, the small patient population recruited into these trials hinders a conclusive assessment on the safety aspects of the CRISPR/Cas9 therapy. Potential safety concerns include the lack of fidelity in the CRISPR/Cas9 system which may lead to unintended DNA modifications at non-targeted gene loci. This review focuses modifications to the CRISPR/ Cas9 components that can mitigate off-target effects in in vitro and preclinical models and its translatability to gene therapy in patient populations.

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Section: Modality Of the Crispr/cas9 System During Deliverymentioning

confidence: 99%

Mitigating off-target effects in CRISPR/Cas9-mediated in vivo gene editing

2020

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“…Gene editing approaches have the advantage of highly specific gene targeting, so unlike LRAs, can produce the desired outcome without global physiological impact. Nevertheless, offtarget effects have been observed in a number of studies and may affect the safety of these methods (Kimberland et al, 2018). So far, the potential of ZFN targeted editing of host CCR5, to induce partial genetic resistance to HIV-1, has been tested in a clinical trial (Tebas et al, 2014).…”

Section: Gene Editingmentioning

confidence: 99%

Measuring the Success of HIV-1 Cure Strategies

Thomas

Ruggiero

Paxton

et al. 2020

Front. Cell. Infect. Microbiol.

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HIV-1 eradication strategies aim to achieve viral remission in the absence of antiretroviral therapy (ART). The development of an HIV-1 cure remains challenging due to the latent reservoir (LR): long-lived CD4 T cells that harbor transcriptionally silent HIV-1 provirus. The LR is stable despite years of suppressive ART and is the source of rebound viremia following therapy interruption. Cure strategies such as "shock and kill" aim to eliminate or reduce the LR by reversing latency, exposing the infected cells to clearance via the immune response or the viral cytopathic effect. Alternative strategies include therapeutic vaccination, which aims to prime the immune response to facilitate control of the virus in the absence of ART. Despite promising advances, these strategies have been unable to significantly reduce the LR or increase the time to viral rebound but have provided invaluable insight in the field of HIV-1 eradication. The development and assessment of an HIV-1 cure requires robust assays that can measure the LR with sufficient sensitivity to detect changes that may occur following treatment. The viral outgrowth assay (VOA) is considered the gold standard method for LR quantification due to its ability to distinguish intact and defective provirus. However, the VOA is time consuming and resource intensive, therefore several alternative assays have been developed to bridge the gap between practicality and accuracy. Whilst a cure for HIV-1 infection remains elusive, recent advances in our understanding of the LR and methods for its eradication have offered renewed hope regarding achieving ART free viral remission.

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“…CRISPR-Cas9 systems has off-targeting potential and inserting the genes between incorrect sequences can lead to very harmful mutations and chromosomal translocations like inducing leukemia (Kimberland, et al 2018). Before the CRISPR systems were prevalent, gene therapies for gene editing even had a higher off target potential and still even with CRISPR mediated systems, reducing the off target effects is of high priority as the patient, who is already in a very weak and worn state will probably not be able to endure those off targeting side effects which were mostly the cases in the gene therapy trials in the past.…”

Section: Where Crispr-cas9 Falls Shortmentioning

confidence: 99%

AIDS tedavisi için gen düzenleme çalışmaları

Akpınaroğlu¹

2020

genediting

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Following after the fatal genetic diseases that were caused by single nucleotide polymorphisms (SNPs) and cancer, HIV/AIDS had always been one of the primary targets of cell and gene therapies due to lack of any proper satisfactory treatment. Earlier gene therapy approaches were mostly trials about introducing anti-HIV genes to cells, using various viral vectors. These viral vectors performed integrations of the desired anti-HIV genes, sometimes correctly while sometimes between random wrong sequences. However, with the increased precision of new gene editing technologies, including ZFNs and the latest CRISPR-mediated gene editing systems (Clustered Regularly Interspaced Short Palindromic Repeats) more successful therapies have begun to be administrated. As an important example of therapy, the trial of Timothy Ray Brown which was followed by the "London patient", allowed the topic of gene editing techniques for treatment of HIV and AIDS to gain interest again. Can Akpinaroglu / Gene Editing Creative Commons Attribution 4.0 International License. www.genediting.net

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Strategies for controlling CRISPR/Cas9 off-target effects and biological variations in mammalian genome editing experiments

Cited by 73 publications

References 108 publications

Mitigating off-target effects in CRISPR/Cas9-mediated in vivo gene editing

Mitigating off-target effects in CRISPR/Cas9-mediated in vivo gene editing

Measuring the Success of HIV-1 Cure Strategies

AIDS tedavisi için gen düzenleme çalışmaları

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