I study delivery technologies for gene therapy, particularly strategies to efficiently deliver the clustered regularly interspaced short palindromic repeats CRISPR/CRISPR-associated (Cas) system for transient expression and efficient genome editing. Transient expression of the editor proteins (e.g., Cas9 protein) is necessary to reduce the risks of mutagenesis from off-target activity and immune responses. I explore the application of bacteriophage RNA-binding proteins (ABPs) and their RNA aptamers in packaging Cas9 mRNA or Cas9 ribonucleoprotein (RNP) into viral capsids.
Our lab successfully developed lentiviral capsid-based bionanoparticles for efficiently delivering CRISPR/Cas9 for transient expression and efficient genome editing. These novel bionanoparticles can deliver Cas9 mRNA or Cas9 RNP. They are highly active in genome editing and have lower off-target rates compared with adeno-associated virus or lentiviral vector delivery. We are further optimizing our delivery system for gene therapy of genetic diseases such as sickle-cell disease and Duchenne muscular dystrophy.
Lyu P, Javidi-Parsijani P, Atala A, Lu B. Delivering Cas9/sgRNA ribonucleoprotein (RNP) by lentiviral capsid-based bionanoparticles for efficient 'hit-and-run' genome editing. Nucleic Acids Res. 2019 Jul 12. pii: gkz605. doi: 10.1093/nar/gkz605. [Epub ahead of print]. PMID: 31299082
Lu B (co-corresponding author), Javidi-Parsijani P*, Makani V* et al. Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing. Nucleic Acids Res. 2019;47(8):e44. doi: 10.1093/nar/gkz093.
Lentiviral-based vectors and related systems and methods for eukaryotic gene editing. Application number: 62/665,080. Filing date: 05/01/2018. Applicants: Baisong Lu, Anthony Atala. (This patent invents a method to package Cas9/sgRNA in lentivirus-like particles for transient expression and efficient gene editing)