RRC ID 52267
著者 Nishimura K, Ohtaka M, Takada H, Kurisaki A, Tran NVK, Tran YTH, Hisatake K, Sano M, Nakanishi M.
タイトル Simple and effective generation of transgene-free induced pluripotent stem cells using an auto-erasable Sendai virus vector responding to microRNA-302.
ジャーナル Stem Cell Res
Abstract Transgene-free induced pluripotent stem cells (iPSCs) are valuable for both basic research and potential clinical applications. We previously reported that a replication-defective and persistent Sendai virus (SeVdp) vector harboring four reprogramming factors (SeVdp-iPS) can efficiently induce generation of transgene-free iPSCs. This vector can express all four factors stably and simultaneously without chromosomal integration and can be eliminated completely from reprogrammed cells by suppressing vector-derived RNA-dependent RNA polymerase. Here, we describe an improved SeVdp-iPS vector (SeVdp(KOSM)302L) that is automatically erased in response to microRNA-302 (miR-302), uniquely expressed in pluripotent stem cells (PSCs). Gene expression and genome replication of the SeVdp-302L vector, which contains miRNA-302a target sequences at the 3' untranslated region of L mRNA, are strongly suppressed in PSCs. Consequently, SeVdp(KOSM)302L induces expression of reprogramming factors in somatic cells, while it is automatically erased from cells successfully reprogrammed to express miR-302. As this vector can reprogram somatic cells into transgene-free iPSCs without the aid of exogenous short interfering RNA (siRNA), the results we present here demonstrate that this vector may become an invaluable tool for the generation of human iPSCs for future clinical applications.
巻・号 23
ページ 13-19
公開日 2017-8-1
DOI 10.1016/j.scr.2017.06.011
PII S1873-5061(17)30111-3
PMID 28666145
MeSH Animals Base Sequence Cell Culture Techniques / methods* Gene Expression Regulation Genetic Vectors / metabolism* Humans Induced Pluripotent Stem Cells / metabolism* Mice MicroRNAs / genetics MicroRNAs / metabolism* Sendai virus / genetics* Transgenes*
IF 4.495
引用数 21
リソース情報
ヒト・動物細胞 EB5(AES0151) 454E2(HPS0077)