球状核酸とCRISPR遺伝子編集という2つの強力な治療技術の組み合わせは、医療治療の展望を変えるかもしれない Combining two powerful therapeutic technologies – Spherical nucleic acids and CRISPR gene editing – may change the landscape of medical therapeutics
2022-10-06 ノースウェスタン大学
今回の研究で、チームは、遺伝子編集に必要なタンパク質Cas9を構造のコアとして使い、その表面にDNAストランドをくっつけて、新しいタイプのSNAを作った。さらに、このSNAに遺伝子編集を行うことができるRNAをあらかじめ搭載し、ペプチドと融合させることで、細胞の区画障壁を移動する能力を制御し、効率を最大化させた。これらのSNAは、他のクラスのSNAと同様に、トランスフェクション剤(遺伝物質を細胞に導入するためにしばしば必要)を使用せずに効果的に細胞に入り、いくつかのヒトおよびマウスの細胞株で32%から47%の高い遺伝子編集効率を示した。
<関連情報>
- https://news.northwestern.edu/stories/2022/10/discovery-broadens-scope-of-use-of-crispr-gene-editing/
- https://pubs.acs.org/doi/10.1021/jacs.2c07913
CRISPR球状核酸 CRISPR Spherical Nucleic Acids
Chi Huang, Zhenyu Han, Michael Evangelopoulos, and Chad A. Mirkin
Journal of the American Chemical Society Published:October 6, 2022
DOI:https://doi.org/10.1021/jacs.2c07913
Abstract
The use of CRISPR/Cas9 systems in genome editing has been limited by the inability to efficiently deliver the key editing components to and across tissues and cell membranes, respectively. Spherical nucleic acids (SNAs) are nanostructures that provide privileged access to both but have yet to be explored as a means of facilitating gene editing. Herein, a new class of CRISPR SNAs are designed and evaluated in the context of genome editing. Specifically, Cas9 ProSNAs comprised of Cas9 cores densely modified with DNA on their exteriors and preloaded with single-guide RNA were synthesized and evaluated for their genome editing capabilities in the context of multiple cell lines. The radial orientation of the DNA on the Cas9 protein surface enhances cellular uptake, without the need for electroporation or transfection agents. In addition, the Cas9 proteins defining the cores of the ProSNAs were fused with GALA peptides on their N-termini and nuclear localization signals on their C-termini to facilitate endosomal escape and maximize nuclear localization and editing efficiency, respectively. These constructs were stable against protease digestion under conditions that fully degrade the Cas9 protein, when not transformed into an SNA, and used to achieve genome editing efficiency between 32 and 47%. Taken together, these novel constructs and advances point toward a way of significantly broadening the scope of use and impact of CRISPR-Cas9 genome editing systems.
