PMID- 27165101
OWN - NLM
STAT- MEDLINE
DCOM- 20170817
LR  - 20171213
IS  - 1572-8781 (Electronic)
IS  - 1387-2176 (Linking)
VI  - 18
IP  - 3
DP  - 2016 Jun
TI  - Incorporation of VSV-G produces fusogenic plasma membrane vesicles capable of 
      efficient transfer of bioactive macromolecules and mitochondria.
PG  - 41
LID - 10.1007/s10544-016-0066-y [doi]
AB  - The objective of this study was to determine if plasma membrane vesicles (PMVs) 
      could be exploited for efficient transfer of macro-biomolecules and mitochondria. 
      PMVs were derived from mechanical extrusion, and made fusogenic (fPMVs) by 
      incorporating the glycoprotein G of vesicular stomatitis virus (VSV-G). Confocal 
      microscopy examination revealed that cytoplasmic proteins and mitochondria were 
      enclosed in PMVs as evidenced by tracing with cytoplasmically localized and 
      mitochondria-targeted EGFP, respectively. However, no fluorescence signal was 
      detected in PMVs from cells whose nucleus was labeled with an EGFP-tagged histone 
      H2B. Consistently, qRT-PCR measurement showed that mRNA, miRNA and mitochondrial 
      DNA decreased slightly; while nuclear DNA was not measureable. Further, Western 
      blot analysis revealed that cytoplasmic and membrane-bound proteins fell 
      inconspicuously while nuclear proteins were barely detecsle. In addition, fPMVs 
      carrying cytoplamic DsRed proteins transduced about ~40 % of recipient cells. The 
      transfer of protein was further confirmed by using the inducible Cre/loxP system. 
      Mitochondria transfer was found in about 20 % recipient cells after incubation 
      with fPMVs for 5 h. To verify the functionalities of transferred mitochondria, 
      mitochodria-deficient HeLa cells (Rho0) were generated and cultivated with fPMVs. 
      Cell enumeration demonstrated that adding fPMVs into culture media stimulated 
      Rho0 cell growth by 100 % as compared to the control. Lastly, MitoTracker and 
      JC-1 staining showed that transferred mitochondria maintained normal shape and 
      membrane potential in Rho0 cells. This study established a time-saving and 
      efficient approach to delivering proteins and mitochondria by using fPMVs, which 
      would be helpful for finding a cure to mitochondria-associated diseases. 
      Graphical abstract Schematic of the delivery of macro-biomolecules and organelles 
      by fPMVs. VSV-G-expressing cells were extruded through a 3 mum polycarbonate 
      membrane filter to generate fusogenic plasma membrane vesicles (fPMVs), which 
      contain bioactive molecules and organelles but not the nucleus. fPMVs can be 
      endocytosed by target cells, while the cargo is released due to low-pH induced 
      membrane fusion. These nucleus-free fPMVs are efficient at delivery of 
      cytoplasmic proteins and mitochondria, leading to recovery of mitochondrial 
      biogenesis and proliferative ability in mitochondria-deficient cells.
FAU - Lin, Hao-Peng
AU  - Lin HP
AD  - Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 
      515063, China.
FAU - Zheng, De-Jin
AU  - Zheng DJ
AD  - Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 
      515063, China.
FAU - Li, Yun-Pan
AU  - Li YP
AD  - Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 
      515063, China.
FAU - Wang, Na
AU  - Wang N
AD  - Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 
      515063, China.
FAU - Chen, Shao-Jun
AU  - Chen SJ
AD  - Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 
      515063, China.
FAU - Fu, Yu-Cai
AU  - Fu YC
AD  - Laboratory of Cell Senescence, Shantou University Medical College, Shantou, 
      Guangdong, 515041, China.
FAU - Xu, Wen-Can
AU  - Xu WC
AD  - Department of Endocrinology, First Affiliated Hospital of Shantou University 
      Medical College, Shantou, Guangdong, 515041, China.
FAU - Wei, Chi-Ju
AU  - Wei CJ
AD  - Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 
      515063, China. chijuwei@stu.edu.cn.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biomed Microdevices
JT  - Biomedical microdevices
JID - 100887374
RN  - 0 (DNA, Mitochondrial)
RN  - 0 (G protein, vesicular stomatitis virus)
RN  - 0 (Histones)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (MicroRNAs)
RN  - 0 (Polycarboxylate Cement)
RN  - 0 (RNA, Messenger)
RN  - 0 (Viral Envelope Proteins)
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - 25766-59-0 (polycarbonate)
SB  - IM
MH  - Cell Line
MH  - Cell Membrane/*metabolism
MH  - Cell Nucleus
MH  - DNA, Mitochondrial/genetics
MH  - Genomics
MH  - Green Fluorescent Proteins/metabolism
MH  - HeLa Cells
MH  - Histones/metabolism
MH  - Humans
MH  - Membrane Glycoproteins/*metabolism
MH  - MicroRNAs/genetics
MH  - Mitochondria/*metabolism
MH  - Polycarboxylate Cement/chemistry
MH  - RNA, Messenger/genetics
MH  - Sequence Analysis, DNA
MH  - Transport Vesicles/*metabolism
MH  - Vesicular stomatitis Indiana virus
MH  - Viral Envelope Proteins/*metabolism
OTO - NOTNLM
OT  - Macro-biomolecules
OT  - Mitochondria
OT  - Plasma membrane vesicles
OT  - Rho0 cells
OT  - VSV-G
EDAT- 2016/05/12 06:00
MHDA- 2017/08/18 06:00
CRDT- 2016/05/12 06:00
PHST- 2016/05/12 06:00 [entrez]
PHST- 2016/05/12 06:00 [pubmed]
PHST- 2017/08/18 06:00 [medline]
AID - 10.1007/s10544-016-0066-y [pii]
AID - 10.1007/s10544-016-0066-y [doi]
PST - ppublish
SO  - Biomed Microdevices. 2016 Jun;18(3):41. doi: 10.1007/s10544-016-0066-y.