Efficient Delivery of CRISPR-Cas9 for Genome Editing via Self-Assembled DNA Nanoclews

By Wujin Sun, Wenyan Ji, Jordan M. Hall, Quanyin Hu, Chao Wang, Chase L. Beisel, Zhen Gu. Angewandte Chemie, 2015

580 citations recorded by the source corpus.

CRISPR-Cas9 represents a promising technology for genome editing, yet means of safe and efficient delivery remain to be fully realized. Here, we report a novel delivery vehicle to deliver the Cas9 protein and single-guide RNA simultaneously based on DNA nanoclews, yarn-like DNA nanoparticles synthesized by rolling circle amplification. The bio-inspired vehicles efficiently loaded Cas9/single-guide RNA complexes and delivered the complexes to the nuclei of human cells, allowing targeted gene disruptions while maintaining cell viability. Editing was most efficient when the DNA nanoclew sequence and the sgRNA guide sequence were partially complementary, offering a design rule for enhancing delivery. Overall, this strategy provides a versatile platform that could be adapted for delivering other DNA-binding proteins or for functional nucleic acids.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/ee7096f1763c2dc447e461a788dcdff9a27af73a

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In Molecular Biology

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Citations in the corpus

• Delivering CRISPR: a review of the challenges and approaches (2018, background): “As an example, Cas9 protein is positively-charged, but oligonucleotides and Cas9:sgRNA RNP are negatively charged (Sun et al., 2015).”
• Engineering the Delivery System for CRISPR-based Genome Editing (2018, background): “Recently the use of nucleic acids themselves as a polymeric substrate for Cas9 RNP delivery has also been explored [52].”
• Nanoparticles in the clinic (2016, background): “While many nanoparticle-based gene therapies are in clinical trials either for gene knockdown or repair, nanoparticle-based gene editing therapies are close behind, although at the moment, these systems are only investigated preclinically.(69,70)”
• Non-viral delivery systems for CRISPR/Cas9-based genome editing: challenges and opportunities (2018, background): “EGFP xenograft tumor models in vivo after intratumoral injection [103].”