کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
7557386 | 1491338 | 2016 | 5 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Knockdown of gene expression by antisense morpholino oligos in preimplantation mouse embryos cultured in vitro
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کلمات کلیدی
FITCPVDFLIFICMOct4DAPIFCSBSA - BSASmall interfering RNA - RNA تداخل کوچکsiRNA - siRNAbovine serum albumin - آلبومین سرم گاوAntisense - آنتیسنسHCG - اچ سی جیInner cell mass - توده سلولی داخلیPreimplantation mouse embryo - جنین جنین قبل از تکاملfetal calf serum - سرم گوساله جنینleukemia inhibitory factor - عامل مهارکننده لوکمیfluorescein isothiocyanate - فلوئورسین ایسوتیوسیاناتKnockdown - نابودیPolyvinylidene fluoride - پلی وینیلیدین فلورایدhuman chorionic gonadotropin - گونادوتروپین کوریونی انسان
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Knockdown of gene expression by antisense morpholino oligos (MOs) is a simple and effective method for analyzing the roles of genes in mammalian cells. Here, we demonstrate the efficient delivery of MOs by Endo-Porter (EP), a special transfection reagent for MOs, into preimplantation mouse embryos cultured in vitro. A fluorescein-labeled control MO was applied for monitoring the incorporation of MOs into developing 2-cell embryos in the presence of varying amounts of EP and bovine serum albumin. In optimized conditions, fluorescence was detected in 2-cell embryos within a 3-h incubation period. In order to analyze the validity of the optimized conditions, an antisense Oct4 MO was applied for knockdown of the synthesis of OCT4 protein in developing embryos from the 2-cell stage. In blastocysts, the antisense Oct4 MO induced a decrease in the amount in OCT4 protein to less than half. An almost complete absence of OCT4-positive cells and nearly complete disappearance of the inner cell mass in the outgrowths of blastocysts were also noted. These phenotypes corresponded with those of Oct4-deficient mouse embryos. Overall, we suggest that the delivery of MOs using EP is useful for the knockdown of gene expression in preimplantation mouse embryos cultured in vitro.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 509, 15 September 2016, Pages 41-45
Journal: Analytical Biochemistry - Volume 509, 15 September 2016, Pages 41-45
نویسندگان
Yuki Sato, Shiori Sato, Takahiro Kikuchi, Asumi Nonaka, Yuki Kumagai, Akira Sasaki, Masayuki Kobayashi,