Intramuscular expression of functional proteins is a promising strategy for therapeutic purposes.Previously,we developed an intramuscular gene delivery method by combining Pluronic L64 and optimized electropulse,which...Intramuscular expression of functional proteins is a promising strategy for therapeutic purposes.Previously,we developed an intramuscular gene delivery method by combining Pluronic L64 and optimized electropulse,which is among the most efficient methods to date.However,plasmid DNAs(pDNAs)in this method were not compressed,making them unstable and inefficient in vivo.We considered that a proper compression of pDNAs by an appropriate material should facilitate gene expression in this L64-electropulse system.Here,we reported our finding of such a material,Epigallocatechin gallate(EGCG),a natural compound in green teas,which could compress and protect pDNAs and significantly increase intramuscular gene expression in the L64-electropulse system.Meanwhile,we found that polyethylenimine(PEI)could also slightly improve exogenous gene expression in the optimal procedure.By analysing the characteristic differences between EGCG and PEI,we concluded that negatively charged materials with strong affinity to nucleic acids and/or other properties suitable for gene delivery,such as EGCG,are better alternatives than cationic materials(like PEI)for muscle-based gene delivery.The results revealed that a critical principle for material/pDNA complex benefitting intramuscular gene delivery/expression is to keep the complex negatively charged.This proof-of-concept study displays the breakthrough in compressing pDNAs and provides a principle and strategy to develop more efficient intramuscular gene delivery systems for therapeutic applications.展开更多
基金the National Natural Science Foundation of China(no.31370972)the 111 Project(no.B16033)the National Key Research and Development Program of China(2017YFB0702600,2017YFB0702603)。
文摘Intramuscular expression of functional proteins is a promising strategy for therapeutic purposes.Previously,we developed an intramuscular gene delivery method by combining Pluronic L64 and optimized electropulse,which is among the most efficient methods to date.However,plasmid DNAs(pDNAs)in this method were not compressed,making them unstable and inefficient in vivo.We considered that a proper compression of pDNAs by an appropriate material should facilitate gene expression in this L64-electropulse system.Here,we reported our finding of such a material,Epigallocatechin gallate(EGCG),a natural compound in green teas,which could compress and protect pDNAs and significantly increase intramuscular gene expression in the L64-electropulse system.Meanwhile,we found that polyethylenimine(PEI)could also slightly improve exogenous gene expression in the optimal procedure.By analysing the characteristic differences between EGCG and PEI,we concluded that negatively charged materials with strong affinity to nucleic acids and/or other properties suitable for gene delivery,such as EGCG,are better alternatives than cationic materials(like PEI)for muscle-based gene delivery.The results revealed that a critical principle for material/pDNA complex benefitting intramuscular gene delivery/expression is to keep the complex negatively charged.This proof-of-concept study displays the breakthrough in compressing pDNAs and provides a principle and strategy to develop more efficient intramuscular gene delivery systems for therapeutic applications.
