近期《自然—方法學(xué)》（Nature Methods）上一項(xiàng)研究得出結(jié)論,，用于目標(biāo)基因組修飾的鋅指核酸酶（ZFN）能夠以蛋白質(zhì)的形態(tài)穿透細(xì)胞膜，這將讓ZFN被投遞至細(xì)胞內(nèi)的過(guò)程變得更簡(jiǎn)單,、更具安全性,。
經(jīng)過(guò)設(shè)計(jì)的核酸酶比如ZFN，能夠讓多個(gè)種類的基因組產(chǎn)生特定變化,，其對(duì)研究和基因療法的輔助作用讓人眼前一亮,。核酸酶一般是被以DNA或RNA的形態(tài)投遞進(jìn)入細(xì)胞內(nèi)，然后產(chǎn)生功能蛋白,。
Carlos Barbas等人報(bào)告稱,，他們?cè)趲追N哺乳動(dòng)物細(xì)胞中觀察到ZFN蛋白自身能夠穿透細(xì)胞膜。當(dāng)ZFN蛋白在細(xì)胞內(nèi)達(dá)到足夠高的濃度時(shí),，便會(huì)產(chǎn)生特定基因組變化,，其效果和ZFN以DNA形態(tài)投遞是一樣的。雖然此前曾認(rèn)為細(xì)胞膜的滲透作用只針對(duì)一小部分類型的蛋白質(zhì)和肽而言,，而未曾有研究證明ZFN也會(huì)用到這種作用,。
和ZFN以DNA形態(tài)投遞相比，ZFN蛋白在目標(biāo)細(xì)胞內(nèi)只會(huì)短暫存在并且其在基因組變化中所產(chǎn)生的脫靶效應(yīng)要低得多,。蛋白投遞可以避免在投遞過(guò)程中由病毒引起的插入突變風(fēng)險(xiǎn)以及由細(xì)胞對(duì)外源DNA的應(yīng)答而引起的細(xì)胞中毒風(fēng)險(xiǎn),。（生物谷Bioon.com）
doi:10.1038/nmeth.2030
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Targeted gene knockout by direct delivery of zinc-finger nuclease proteins
Thomas Gaj, Jing Guo, Yoshio Kato, Shannon J Sirk & Carlos F Barbas III
Zinc-finger nucleases (ZFNs) are versatile reagents that have redefined genome engineering. Realizing the full potential of this technology requires the development of safe and effective methods for delivering ZFNs into cells. We demonstrate the intrinsic cell-penetrating capabilities of the standard ZFN architecture and show that direct delivery of ZFNs as proteins leads to efficient endogenous gene disruption in various mammalian cell types with minimal off-target effects.
