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Science采用改进CRISPR技术 发现近500个新的lncRNAs

2016-12-20 生物通/万纹 生物通/万纹

长链非编码RNA (lncRNAs)是一类长度大约为200个核苷酸,但不编码任何蛋白的神秘分子,一直以来科学家们都想知道基因组中这类分子到底是起什么作用的。12月15日Science杂志公布了一项最新研究发现:来自加州大学旧金山分校的研究人员识别出了499个新lncRNAs,这为理解这些小分子的功能奠定了基础。同时研究人员也发现lncRNAs与大多数编码基因不同,后者能作用于多种细胞系,但令人惊讶


长链非编码RNA (lncRNAs)是一类长度大约为200个核苷酸,但不编码任何蛋白的神秘分子,一直以来科学家们都想知道基因组中这类分子到底是起什么作用的。

12月15日Science杂志公布了一项最新研究发现:来自加州大学旧金山分校的研究人员识别出了499个新lncRNAs,这为理解这些小分子的功能奠定了基础。同时研究人员也发现lncRNAs与大多数编码基因不同,后者能作用于多种细胞系,但令人惊讶的是,90%已发现的lncRNAs只会影响一种细胞类型的生长。

“大多数细胞中都具有大量的lncRNAs这种神秘分子,但我们不知道它们具体有多少,哪些对基本生物功能具有重要意义。这项最新成果让我们惊讶的是,大多数lncRNAs只在一种细胞类型内发挥功能,这种RNA功能的极度特异性是之前未曾发现过的,”文章作者,加州大学旧金山分校神经学家和临床医师 Daniel Lim说。

之前的研究表明,某些lncRNAs 在细胞生长和功能,以及疾病发生过程中扮演了关键角色,十年前科学家们发现人体基因组中很大一部分是转录成了lncRNAs,一些研究组指出几十个lncRNAs对细胞,甚至机体来说具有重要意义。但只有少数研究发现了lncRNAs的功能,这受限于分析lncRNAs功能和作用细胞类型的研究工具。

在最新这项研究中,研究人员采用了非常热门的一项新工具:CRISPR技术,研发了一种CRISPR干扰平台(CRISPRi),靶向七种不同细胞系中16,401个lncRNAs,这七种细胞系包括六种转化细胞系,和一种人诱导多能干细胞系。研究人员利用CRISPRi工具,抑制了每种候选lncRNA,这样筛选了上千个位点,从中找到了499个细胞强健生长所必需的位点,而其中89%只作用于一种细胞类型。

“这种技术的力量来自CRISPRi,是它帮助我们能关闭这些非编码转录子的转录,”Lim说。为了能确保CRISPRi快速筛选顺利进行,研究人员也为每个位点准备了10个导向RNA,从而构建了大约包含170,000个导向RNAs的文库,研究人员将这个文库命名为 CRiNCL (CRISPRi noncoding library),由Addgene提供。

不过这项研究中最令人惊奇的发现,就是绝大多数的lncRNA基因只作用于一种细胞类型的细胞生长。蛋白编码基因在许多类型细胞中发挥着关键的作用,但lncRNAs只在一种细胞中扮演重要角色,这说明lncRNAs给每个细胞注明了身份。

一些科学家对这一研究盛赞,指出这是第一次通过特殊细胞类型全基因组筛选lncRNA,从中我们可以断定大多数lncRNAs对于细胞活力至关重要,之前这一观点还被质疑,现在得到了证实。

不过目前尚不清楚这499个lncRNAs位点如何影响细胞生长,这一研究克服了全基因组筛选的许多不足之处,但不能判断到底是这些lncRNAs转录的作用,还是最终RNA产物的作用。

同时这项研究也将对靶向癌症治疗产生影响,研究证明了许多lncRNAs位点在细胞增殖过程中具有重要意义,那么在癌症等需要细胞增殖的疾病中,这些位点就代表着以RNA为基础的新型治疗途径。

原始出处

S. John Liu1,2,*, Max A. Horlbeck3,4,5,6,*, Seung Woo Cho9.et.al.CRISPRi-based genome-scale identification of functional long noncoding RNA loci in human cells.Science  15 Dec 2016

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    2017-03-10 dzx0922889
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    2016-12-22 yuandd
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    2016-12-22 xzw113
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    2016-12-22 jichang
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