Centre for Molecular Medicine and Therapeutics at British Columbia Children's Hospital, The University of British Columbia, Vancouver, BC, Canada; Department of Medical Genetics, The University of British Columbia, Vancouver, BC, Canada.
Centre for Molecular Medicine and Therapeutics at British Columbia Children's Hospital, The University of British Columbia, Vancouver, BC, Canada.
J Control Release. 2022 Oct;350:401-413. doi: 10.1016/j.jconrel.2022.08.042. Epub 2022 Aug 27.
CRISPR/Cas9-based genome-editing therapies are poised to change the clinical outcome for many diseases with validated therapeutic targets awaiting an appropriate delivery system. Recent advances in lipid nanoparticle (LNP) technology make them an attractive platform for the delivery of various forms of CRISPR/Cas9, including the efficient and transient Cas9/gRNA ribonucleoprotein (RNP) complexes. In this study, we initially tested our novel LNP platform by delivering pre-complexed RNPs and template DNA to cultured mouse cortical neurons, and obtained successful ex vivo genome editing. We then directly injected LNP-packaged RNPs and DNA template into the mouse cornea to evaluate in vivo delivery. For the first time, we demonstrated wide-spread genome editing in the cornea using our LNP-RNPs. The ability of our LNPs to transfect the cornea highlights the potential of our novel delivery platform to be used in CRISPR/Cas9-based genome editing therapies of corneal diseases.
基于 CRISPR/Cas9 的基因组编辑疗法有望改变许多疾病的临床疗效，因为有许多经过验证的治疗靶点等待合适的递送系统。最近脂质纳米颗粒（LNP）技术的进步使它们成为递送各种形式的 CRISPR/Cas9 的有吸引力的平台，包括高效和瞬时的 Cas9/gRNA 核糖核蛋白（RNP）复合物。在这项研究中，我们最初通过向培养的小鼠皮质神经元递送预复合物的 RNP 和模板 DNA 来测试我们的新型 LNP 平台，并获得了成功的离体基因组编辑。然后，我们直接将 LNP 包裹的 RNP 和 DNA 模板注射到小鼠角膜中，以评估体内递送。我们首次使用我们的 LNP-RNPs 在角膜中实现了广泛的基因组编辑。我们的 LNPs 转染角膜的能力突出了我们的新型递送平台在基于 CRISPR/Cas9 的角膜疾病基因组编辑疗法中的应用潜力。
