Nanoparticle delivery of Cas9 ribonucleoprotein and donor DNA in vivo induces homology-directed DNA repair.
Kunwoo LeeMichael ConboyHyo Min ParkFuguo JiangHyun Jin KimMark A DewittVanessa A MackleyKevin ChangAnirudh RaoColin SkinnerTamanna ShobhaMelod MehdipourHui LiuWen-Chin HuangFreeman LanNicolas L BraySong LiJacob E CornKazunori KataokaJennifer A DoudnaIrina ConboyNiren MurthyPublished in: Nature biomedical engineering (2017)
CRISPR/Cas9-based therapeutics, especially those that can correct gene mutations via homology directed repair (HDR), have the potential to revolutionize the treatment of genetic diseases. However, HDR-based therapeutics are challenging to develop because they require simultaneous in vivo delivery of Cas9 protein, guide RNA and donor DNA. Here, we demonstrate that a delivery vehicle composed of gold nanoparticles conjugated to DNA and complexed with cationic endosomal disruptive polymers can deliver Cas9 ribonucleoprotein and donor DNA into a wide variety of cell types, and efficiently correct the DNA mutation that causes Duchenne muscular dystrophy in mice via local injection, with minimal off-target DNA damage.
Keyphrases
- crispr cas
- circulating tumor
- genome editing
- dna damage
- dna repair
- cell free
- single molecule
- duchenne muscular dystrophy
- gold nanoparticles
- nucleic acid
- gene expression
- oxidative stress
- photodynamic therapy
- single cell
- cell therapy
- metabolic syndrome
- combination therapy
- replacement therapy
- insulin resistance
- dna damage response
- climate change