發(fā)布時(shí)間：2018-04-11 08:29

  本文選題：syncytinA + 條件敲除小鼠�。� 參考：《實(shí)驗(yàn)動(dòng)物科學(xué)》2015年05期

【摘要】：目的構(gòu)建syncytinA(合胞素A)條件敲除小鼠,為進(jìn)一步研究syncytinA在胎盤形成過程中發(fā)揮的融合及非融合作用及研究子癇前期病理模型提供基礎(chǔ)。方法在用ES細(xì)胞打靶完成syncytinA外顯子上游loxp同源重組基礎(chǔ)上,利用CRISPR-Cas9得到syncytinA-loxp小鼠。構(gòu)建syncytinA-loxp轉(zhuǎn)基因載體及sg RNA,通過原核顯微注射方法將構(gòu)建好的doner、Cas9及sg RNA一并注射到C57小鼠受精卵中,并移植入同期受孕代孕受體ICR輸卵管中獲得子代小鼠。用PCR方法檢測子代鼠尾基因型,loxp陽性小鼠與WT交配獲得syncytinA-loxp小鼠。為了檢測syncytinA-loxp能否被敲除,通過與prime1cre及zp3cre交配獲得syncytinA-/-,PCR及q-PCR檢測syncytinA是否被敲掉。結(jié)果經(jīng)PCR及q-PCR方法檢測,我們成功得到syncytinA-/-胚胎。結(jié)論 syncytinA條件敲除小鼠構(gòu)建成功,為更好的研究它在胎盤及子癇前期中的功能提供了基礎(chǔ)。
[Abstract]:Objective to construct syncytin A (syncytin A) conditioned knockout mice and to provide a basis for further study on the fusion and non-fusion role of syncytinA in placental formation and the study of preeclampsia pathological model.Methods on the basis of loxp homologous recombination in upstream of syncytinA exon with es cells, syncytinA-loxp mice were obtained by CRISPR-Cas9.SyncytinA-loxp transgenic vectors and sg RNAs were constructed. The constructed donercas 9 and sg RNA were injected into the fertilized eggs of C57 mice by prokaryotic microinjection, and then transplanted into the oviduct of the surrogate ICR receptor to obtain the offspring mice.SyncytinA-loxp mice were obtained by mating with WT by PCR method.In order to detect whether syncytinA-loxp can be knocked out, syncytin A / P -PCR and q-PCR were used to detect whether syncytinA was knocked out by mating with prime1cre and zp3cre.Results by PCR and q-PCR methods, we successfully obtained syncytin A-r-embryos.Conclusion the successful construction of syncytinA conditioned knockout mice provides a basis for better study of its function in placenta and preeclampsia.
【作者單位】： 首都醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院醫(yī)學(xué)遺傳與發(fā)育生物學(xué)系;北京生命科學(xué)研究所轉(zhuǎn)基因動(dòng)物中心;基礎(chǔ)醫(yī)學(xué)系默索大學(xué)醫(yī)學(xué)院紀(jì)念健康醫(yī)學(xué)中心婦科與婦產(chǎn)科系;
【基金】：北京市自然科學(xué)基金(No.7142026)
【分類號】：R-332;R714.244
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本文編號：1735208