Method and reagent for preventing and treating heart disease
A heart disease, inhibitor technology, applied in cardiovascular system diseases, biochemical equipment and methods, gene therapy, etc., can solve problems such as unproven miRNA
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[0062] The preparation method of the small interfering RNA is not particularly limited in the present invention, including but not limited to: chemical synthesis method, in vitro transcription method and the like. It should be understood that those skilled in the art can conveniently prepare or express the small interfering RNA in various ways after knowing the sequence of the siRNA provided by the present invention. For example, in a preferred embodiment of the present invention, the small interfering RNA is chemically synthesized.
[0063] Small interfering RNA can be prepared in the form of double-stranded nucleic acid, which contains a sense strand and an antisense strand, and these two strands form a double strand only under hybridization conditions. A double-stranded RNA complex can be prepared from separate sense and antisense strands. Thus, for example, complementary sense and antisense strands are chemically synthesized and subsequently hybridized by annealing to pro...
Embodiment 1
[0120] Embodiment 1, human heart miRNA expression profiling analysis
[0121] Regarding which miRNA is most abundantly expressed in the heart, the results of previous studies in different laboratories are different. Therefore, the present inventors used the small RNA chip method to analyze the expression profile of miRNAs in human cardiac total RNA. Of the 718 human miRNAs detected, only 236 miRNAs were detected. Among the detected miRNAs, 41 miRNAs with a signal value above 1000 were expressed in a high abundance in the heart. figure 1 23 miRNAs with signal values higher than 4000 are shown. Among them, miR-1 has the highest signal value, indicating that miR-1 is the miRNA with the highest expression in the heart. Several miRNA families including let-7, miR-26, miR-126 and miR-133 were also expressed in higher abundance.
Embodiment 2
[0122] Example 2, Heart-enriched miRNA identification and cardiac miRNA sequence terminal heterogeneity
[0123] To verify heart-enriched miRNAs, the inventors constructed a small RNA library using human heart total RNA, and sequenced the obtained small RNA clones. Among the 251 clones sequenced, 113 (45%) were known miRNA sequences, and the remaining small RNA clones were rRNA, tRNA, mRNA and human genome repeat fragments (data not shown). In miRNA cloning, the inventors identified 27 different miRNAs, of which 15 were cloned multiple times and the remaining 12 were cloned only once. Most of the miRNAs (22 / 27) obtained by cloning are in the high-abundance miRNA list of the chip data results above, indicating the consistency of the results obtained by the two methods. Importantly, the number of clones of miR-1 accounts for 24% (27 / 113) of the total number of miRNA clones, indicating that miR-1 is indeed the most abundant miRNA in the heart at least in the inventor's study.
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