本文選題：豬　切入點(diǎn)：ZBED基因　出處：《中國畜牧雜志》2017年08期 　論文類型：期刊論文

【摘要】：鋅指蛋白ZBED6基因的突變可促進(jìn)骨骼肌中胰島素樣生長因子2(IGF2)轉(zhuǎn)錄水平顯著上調(diào),從而促進(jìn)細(xì)胞增殖和肌管形成。本研究利用CRISPR-Cas9技術(shù)對(duì)豬的ZBED6基因進(jìn)行編輯,利用在線軟件篩選出4個(gè)突變靶點(diǎn),利用p Cas9/g RNA載體構(gòu)建p Cas9/g RNA-ZBED6質(zhì)粒,再構(gòu)建4個(gè)用于驗(yàn)證突變效率的p TYNE-ZBED6質(zhì)粒,同時(shí)轉(zhuǎn)染HEK293細(xì)胞系,通過觀察熒光信號(hào)的強(qiáng)弱判斷靶序列突變效率的高低。結(jié)果表明:位于編碼區(qū)第17~37位點(diǎn)的靶點(diǎn)1轉(zhuǎn)染后熒光信號(hào)最強(qiáng),切割效率最高,該位點(diǎn)是利用CRISPR-Cas9技術(shù)編輯ZBED6基因的有效位點(diǎn)。
[Abstract]:The mutation of zinc finger protein ZBED6 gene can significantly up-regulate the transcription level of insulin-like growth factor-2 IGF2 in skeletal muscle, thus promote cell proliferation and myotube formation. In this study, CRISPR-Cas9 technique was used to edit porcine ZBED6 gene. Four mutation targets were screened out by on-line software, then p Cas9/g RNA-ZBED6 plasmid was constructed by using p Cas9/g RNA vector, and four p TYNE-ZBED6 plasmids were constructed to verify the mutation efficiency. At the same time, they were transfected into HEK293 cell line. The mutation efficiency of target sequence was judged by observing the intensity of fluorescence signal. The results showed that target 1, located at site 1737 of coding region, had the strongest fluorescence signal and the highest cutting efficiency after transfection. This site is an effective site for editing ZBED6 gene by CRISPR-Cas9 technique.
【作者單位】： 黑龍江省農(nóng)業(yè)科學(xué)院畜牧研究所;農(nóng)業(yè)部畜禽遺傳資源與種質(zhì)創(chuàng)新重點(diǎn)實(shí)驗(yàn)室;
【基金】：農(nóng)業(yè)部畜禽遺傳資源與種質(zhì)創(chuàng)新重點(diǎn)實(shí)驗(yàn)室開放課題 國家生豬產(chǎn)業(yè)技術(shù)體系(CARS-36)
【分類號(hào)】：Q78;S828

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