RRC ID 48006
Author Shoji K, Suzuki Y, Sugano S, Shimada T, Katsuma S.
Title Artificial "ping-pong" cascade of PIWI-interacting RNA in silkworm cells.
Journal RNA
Abstract PIWI-interacting RNAs (piRNAs) play essential roles in the defense system against selfish elements in animal germline cells by cooperating with PIWI proteins. A subset of piRNAs is predicted to be generated via the "ping-pong" cascade, which is mainly controlled by two different PIWI proteins. Here we established a cell-based artificial piRNA production system using a silkworm ovarian cultured cell line that is believed to possess a complete piRNA pathway. In addition, we took advantage of a unique silkworm sex-determining one-to-one ping-pong piRNA pair, which enabled us to precisely monitor the behavior of individual artificial piRNAs. With this novel strategy, we successfully generated artificial piRNAs against endogenous protein-coding genes via the expected back-and-forth traveling mechanism. Furthermore, we detected "primary" piRNAs from the upstream region of the artificial "ping-pong" site in the endogenous gene. This artificial piRNA production system experimentally confirms the existence of the "ping-pong" cascade of piRNAs. Also, this system will enable us to identify the factors involved in both, or each, of the "ping" and "pong" cascades and the sequence features that are required for efficient piRNA production.
Volume 23(1)
Pages 86-97
Published 2017-1-1
DOI 10.1261/rna.058875.116
PII rna.058875.116
PMID 27777367
PMC PMC5159652
MeSH Animals Bombyx / genetics* Cell Line Cells, Cultured Female Insect Proteins / genetics Insect Proteins / metabolism Ovary / cytology* Ovary / metabolism RNA, Small Interfering / genetics*
IF 4.32
Times Cited 4