Short interfering RNA-induced gene silencing is transmitted between cells from the mammalian central nervous system.
Although short interfering RNA (siRNA)-induced gene silencing can be transmitted between cells in plants and in Caenorhabditis elegans, this phenomenon has been barely studied in mammalian cells. Both immortalized oligodendrocytes and SNB19 glioblastoma cells were transfected with siRNA constructs for phosphatase and tensin homolog deleted on chromosome 10 (PTEN) or Akt/protein kinase B (Akt). Co-cultures were established between silenced cells and non-silenced cells which were hygromycin resistant and/or expressed green fluorescent protein. After fluorescence sorting or hygromycin selection to remove the silenced cells, the expression of PTEN or Akt genes in the originally unsilenced cells was in all cases significantly decreased. Importantly, silencing did not occur in transwell culture studies, suggesting that transmission of the silencing signal requires a close association between cells. These results provide the first direct demonstration that an siRNA-induced silencing signal can be transmitted between mammalian CNS cells.
Zhao
Tian-Yong
TY
Department of Anatomy and Neurobiology, University of Kentucky College of Medicine, Lexington, Kentucky 40536-0298, USA.
Zou
Shi-Ping
SP
Alimova
Yelena V
YV
Wang
Guoying
G
Hauser
Kurt F
KF
Ghandour
M Said
MS
Knapp
Pamela E
PE
D015152:Blotting, Northern / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D002490:Central Nervous System / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D002875:Chromosomes / Q000378:metabolism; D018920:Coculture Techniques / Q000379:methods; D017353:Gene Deletion; D015870:Gene Expression / Q000187:drug effects*; D020868:Gene Silencing / Q000187:drug effects*; D022761:Gene Transfer, Horizontal / Q000187:drug effects*; D005909:Glioblastoma; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D009836:Oligodendroglia; D051059:PTEN Phosphohydrolase / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D034741:RNA, Small Interfering / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014162:Transfection / Q000379:methods
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D051059:PTEN Phosphohydrolase