高效能沉默BMP-7基因siRNA序列的實驗篩選
發(fā)布時間：2018-10-11 20:07
【摘要】：目的骨形態(tài)發(fā)生蛋白(bone morphogenetic protein-7,BMP-7)是一種促進骨和軟骨再生的關(guān)鍵性生長因子,旨在篩選高效能BMP-7si RNA序列,為后續(xù)探討B(tài)MP-7的生物學(xué)作用機制的基礎(chǔ)和臨床應(yīng)用研究提供實驗研究技術(shù)手段。方法本實驗針對BMP-7基因,構(gòu)建出相應(yīng)4條si RNA序列,分別將其轉(zhuǎn)染至大鼠臍靜脈內(nèi)皮細胞,轉(zhuǎn)染72 h后用Wb和RT-PCR方法檢測BMP-7蛋白和BMP-7 m RNA表達水平,篩選出最高效的BMP-7si RNA序列。結(jié)果人工合成的4條BMP-7si RNA序列都能有效抑制大鼠臍靜脈內(nèi)皮細胞的BMP-7表達,其中BMP-7-3si RNA序列抑制效果最為明顯,抑制率大于90%,被確認為體外抑制BMP-7表達最為有效的干擾序列。結(jié)論 BMP-7-3 si RNA序列能夠在體外高效能抑制BMP-7表達,故可作為一種實驗研究技術(shù)手段,用于探討B(tài)MP-7的生物學(xué)作用機制的基礎(chǔ)和臨床應(yīng)用研究。
[Abstract]:Objective Bone morphogenetic protein (bone morphogenetic protein-7,BMP-7) is a key growth factor to promote bone and cartilage regeneration. To provide experimental techniques for the further study of the biological mechanism and clinical application of BMP-7. Methods four si RNA sequences of BMP-7 gene were constructed and transfected into rat umbilical vein endothelial cells. After 72 h of transfection, the expression levels of BMP-7 protein and BMP-7 m RNA were detected by Wb and RT-PCR methods, and the most efficient BMP-7si RNA sequences were screened out. Results all of the four synthetic BMP-7si RNA sequences could effectively inhibit the expression of BMP-7 in rat umbilical vein endothelial cells. The inhibitory rate of BMP-7-3si RNA sequence was more than 90, which was confirmed as the most effective interference sequence to inhibit the expression of BMP-7 in vitro. Conclusion BMP-7-3 si RNA sequence can effectively inhibit the expression of BMP-7 in vitro, so it can be used as an experimental technique to study the biological mechanism and clinical application of BMP-7.
【作者單位】： 昆明醫(yī)科大學(xué);昆明醫(yī)科大學(xué)第一附屬醫(yī)院;
【基金】：國家自然科學(xué)基金(81160239,81460296) 云南省科技廳-昆明醫(yī)科大學(xué)聯(lián)合專項重點項目(2012FB161)~~
【分類號】：R3416
[Abstract]:Objective Bone morphogenetic protein (bone morphogenetic protein-7,BMP-7) is a key growth factor to promote bone and cartilage regeneration. To provide experimental techniques for the further study of the biological mechanism and clinical application of BMP-7. Methods four si RNA sequences of BMP-7 gene were constructed and transfected into rat umbilical vein endothelial cells. After 72 h of transfection, the expression levels of BMP-7 protein and BMP-7 m RNA were detected by Wb and RT-PCR methods, and the most efficient BMP-7si RNA sequences were screened out. Results all of the four synthetic BMP-7si RNA sequences could effectively inhibit the expression of BMP-7 in rat umbilical vein endothelial cells. The inhibitory rate of BMP-7-3si RNA sequence was more than 90, which was confirmed as the most effective interference sequence to inhibit the expression of BMP-7 in vitro. Conclusion BMP-7-3 si RNA sequence can effectively inhibit the expression of BMP-7 in vitro, so it can be used as an experimental technique to study the biological mechanism and clinical application of BMP-7.
【作者單位】： 昆明醫(yī)科大學(xué);昆明醫(yī)科大學(xué)第一附屬醫(yī)院;
【基金】：國家自然科學(xué)基金(81160239,81460296) 云南省科技廳-昆明醫(yī)科大學(xué)聯(lián)合專項重點項目(2012FB161)~~
【分類號】：R3416
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