: K7 C m& M7 p* l目前研究人员已经得到的miRNA开关包括miR-1-,miR-208a-,miR499a-5p-等，这几种miRNA开关均能够有效纯化从人类多能干细胞诱导分化而来的心肌细胞。并且不同的miRNA开关在细胞发育不同阶段作用效果不同，这表明可以根据心肌细胞发育不同阶段表达的不同miRNA将心肌细胞进行分段纯化筛选。 u) r+ R/ }0 I6 m# x- k
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研究人员相信，在未来的研究中，miRNA开关可以对不同类型的和不同发育阶段的细胞进行纯化筛选，对于细胞生物学和发育生物学研究具有重要意义。（生物谷Bioon.com） 0 W- B* y! W3 ]6 t5 q' U 7 n4 \6 ^6 w/ |( s$ q ( B) K3 Y7 k& m/ }8 V- ^ $ c. P1 S* v% {6 N. B+ D& V0 S* n$ G% c. @* G 9 l0 s6 ~# z4 q* y& ~" H
/ d+ Y5 [8 ^1 o5 g; `) Y( }% I DOI: [url]http://dx.doi.org/10.1016/j.stem.2015.04.005. ~1 f9 u# F# r2 X% m8 U6 J
/ z8 R9 }* t1 n, |Efficient Detection and Purification of Cell Populations Using Synthetic MicroRNA Switches 4 N) m% Y: V2 N5 C; n- { 9 T# h3 }1 l/ l8 R- W2 u' zKenji Miki7, Kei Endo7, Seiya Takahashi, Shunsuke Funakoshi, Ikue Takei, Shota Katayama, Taro Toyoda, Maki Kotaka, Tadashi Takaki, Masayuki Umeda, Chikako Okubo, Misato Nishikawa, Akiko Oishi, Megumi Narita, Ito Miyashita, Kanako Asano, Karin Hayashi, Kenji Osafune, Shinya Yamanaka, Hirohide Saitocorrespondenceemail, Yoshinori Yoshida ) y# ]: C) l0 [& G6 E. B4 c) u. L$ B1 ?- l- u
Isolation of specific cell types, including pluripotent stem cell (PSC)-derived populations, is frequently accomplished using cell surface antigens expressed by the cells of interest. However, specific antigens for many cell types have not been identified, making their isolation difficult. Here, we describe an efficient method for purifying cells based on endogenous miRNA activity. We designed synthetic mRNAs encoding a fluorescent protein tagged with sequences targeted by miRNAs expressed by the cells of interest. These miRNA switches control their translation levels by sensing miRNA activities. Several miRNA switches (miR-1-, miR-208a-, and miR-499a-5p-switches) efficiently purified cardiomyocytes differentiated from human PSCs, and switches encoding the apoptosis inducer Bim enriched for cardiomyocytes without cell sorting. This approach is generally applicable, as miR-126-, miR-122-5p-, and miR-375-switches purified endothelial cells, hepatocytes, and insulin-producing cells differentiated from hPSCs, respectively. Thus, miRNA switches can purify cell populations for which other isolation strategies are unavailable. 1 c! [4 z' |! E0 G& N2 }9 @$ X; @! a& o% n/ O( z, m
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2楼原文 感谢Damon-Salvatore 提供作者: Damon-Salvatore 时间: 2015-5-27 15:44
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