通过沃森-克里克理论识别单链的γ肽核酸的体内特定位点的基因编辑

Title:Single-Stranded γPNAs for In Vivo Site-Specific Genome Editing via Watson-Crick Recognition

Volume: 14 Issue: 5

Author(s): Raman Bahal, Elias Quijano, Nicole A. McNeer, Yanfeng Liu, Dinesh C. Bhunia, Francesco Lopez-Giraldez, Rachel J. Fields, William M. Saltzman, Danith H. Ly and Peter M. Glazer

Affiliation:

关键词: β-球蛋白，基因组编辑，绿色荧光蛋白，纳米颗粒，聚乳酸-羟基乙酸共聚物，肽核酸。

摘要: Triplex-forming peptide nucleic acids (PNAs) facilitate gene editing by stimulating recombination of donor DNAs within genomic DNA via site-specific formation of altered helical structures that further stimulate DNA repair. However, PNAs designed for triplex formation are sequence restricted to homopurine sites. Herein we describe a novel strategy where next generation single-stranded gamma PNAs (γPNAs) containing miniPEG substitutions at the gamma position can target genomic DNA in mouse bone marrow at mixed-sequence sites to induce targeted gene editing. In addition to enhanced binding, γPNAs confer increased solubility and improved formulation into poly(lactic-co-glycolic acid) (PLGA) nanoparticles for efficient intracellular delivery. Single-stranded γPNAs induce targeted gene editing at frequencies of 0.8% in mouse bone marrow cells treated ex vivo and 0.1% in vivo via IV injection, without detectable toxicity. These results suggest that γPNAs may provide a new tool for induced gene editing based on Watson-Crick recognition without sequence restriction.

Cite this article as:

Bahal Raman, Quijano Elias, McNeer A. Nicole, Liu Yanfeng, Bhunia C. Dinesh, Lopez-Giraldez Francesco, Fields J. Rachel, Saltzman M. William, Ly H. Danith and Glazer M. Peter, Single-Stranded γPNAs for In Vivo Site-Specific Genome Editing via Watson-Crick Recognition, Current Gene Therapy 2014; 14 (5) . https://dx.doi.org/10.2174/1566523214666140825154158