Reduced off-target effect of NG-BE4max by using NG-HiFi system

Shan, Huanhuan; Liu, Zhiquan; Jia, Yingqi; Chen, Siyu; Chen, Mao; Song, Yuning; Sui, Tingting; Lai, Liangxue; Li, Zhanjun

doi:10.1016/j.omtn.2021.05.012

Cited by 3 publications

(1 citation statement)

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“…82 To correct TGM1 c.877−2 A > G, we selected the cytosine base editor NG-BE4max, which targets the noncoding strand, thus, causing a base exchange from the mutant G back to the wild-type A on our target strand. 83,84 We then prepared modified and unmodified base editor mRNA and assessed their editing efficacy in ARCI patient cells using RNAimax. This treatment resulted in editing rates of 4.6 ± 0.9% with the modified mRNA, while unmodified (uridine 5′-triphosphate) mRNA yielded no detectable edits (Figure 7C).…”

Section: Lnp Composition and Genetic Payload Determinementioning

confidence: 99%

Lipid Nanoparticle-Mediated Hit-and-Run Approaches Yield Efficient and Safe In Situ Gene Editing in Human Skin

Bolsoni,

Liu,

Mohabatpour

et al. 2023

ACS Nano

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Despite exciting advances in gene editing, the efficient delivery of genetic tools to extrahepatic tissues remains challenging. This holds particularly true for the skin, which poses a highly restrictive delivery barrier. In this study, we ran a head-to-head comparison between Cas9 mRNA or ribonucleoprotein (RNP)-loaded lipid nanoparticles (LNPs) to deliver gene editing tools into epidermal layers of human skin, aiming for in situ gene editing. We observed distinct LNP composition and cell-specific effects such as an extended presence of RNP in slow-cycling epithelial cells for up to 72 h. While obtaining similar gene editing rates using Cas9 RNP and mRNA with MC3-based LNPs (10−16%), mRNA-loaded LNPs proved to be more cytotoxic. Interestingly, ionizable lipids with a pK a ∼ 7.1 yielded superior gene editing rates (55%−72%) in two-dimensional (2D) epithelial cells while no single guide RNA-dependent off-target effects were detectable. Unexpectedly, these high 2D editing efficacies did not translate to actual skin tissue where overall gene editing rates between 5%−12% were achieved after a single application and irrespective of the LNP composition. Finally, we successfully base-corrected a disease-causing mutation with an efficacy of ∼5% in autosomal recessive congenital ichthyosis patient cells, showcasing the potential of this strategy for the treatment of monogenic skin diseases. Taken together, this study demonstrates the feasibility of an in situ correction of disease-causing mutations in the skin that could provide effective treatment and potentially even a cure for rare, monogenic, and common skin diseases.

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Section: Lnp Composition and Genetic Payload Determinementioning

confidence: 99%