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 develop lentivirus-like particles, exosomes and nanoparticles for efficient and safe delivery of Cas9 mRNA or Cas9 ribonucleoprotein (RNP) in vitro and in vivo.
CRISPR/Cas9 can generate large on target DNA deletions which may affect safety and efficiency, I fused DNA polymerase to Cas9 and successfully reduced deletion size and improved gene editing efficiency in primary human cells.
I am also developing gene therapy for type I Dent’s disease, a rare genetic disease caused by mutations in CLCN5 gene and affecting the kidney function. I use lentiviral vectors to deliver functional CLCN5 cDNA to the kidney of disease animal models and see great therapeutic effects. Achieving sustained therapeutic effects is My next goal.
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)