A droplet digital PCR detection method for rare L1 insertions in tumors.
A droplet digital PCR detection method for rare L1 insertions in tumors.
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DOI：
10.1186/s13100-014-0030-4
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发表时间：
2014
期刊：
影响因子：
4.9
通讯作者：
Deininger PL
中科院分区：
文献类型：
--
作者：
White TB;McCoy AM;Streva VA;Fenrich J;Deininger PL
活跃的人类移动元件，长散布元件1 （L1）目前在每个单倍体基因组中超过50万拷贝。通过其通过靶引反转录（TPRT）过程的迁移，L1的动员已导致超过100例人类疾病的新发病例，并且最近与各种癌症类型相关。高通量测序（HTS）技术的巨大进步使人们对L1在人类癌症中的作用有了更多的了解；然而，研究人员仍然受到验证HTS检测到的潜在罕见L1插入事件的能力的限制，这些事件可能只发生在一小部分肿瘤细胞中。此外，HTS对稀有事件的检测随着读取深度的变化而变化很大，需要新的元素发现工具来填补HTS造成的空白。我们采用液滴数字PCR （ddPCR）检测了人类马赛克基因组中罕见的L1位点。我们的检测方法可以检测到每10000个细胞中只有一个细胞的L1插入。ddPCR是一种可靠的方法，可与HTS技术一起用于检测、验证和定量肿瘤和其他组织中罕见的L1插入事件。本文的在线版本（doi:10.1186/s13100-014-0030-4）包含补充资料，可供授权用户使用。
The active human mobile element, long interspersed element 1 (L1) currently populates human genomes in excess of 500,000 copies per haploid genome. Through its mobility via a process called target primed reverse transcription (TPRT), L1 mobilization has resulted in over 100 de novo cases of human disease and has recently been associated with various cancer types. Large advances in high-throughput sequencing (HTS) technology have allowed for an increased understanding of the role of L1 in human cancer; however, researchers are still limited by the ability to validate potentially rare L1 insertion events detected by HTS that may occur in only a small fraction of tumor cells. Additionally, HTS detection of rare events varies greatly as a function of read depth, and new tools for de novo element discovery are needed to fill in gaps created by HTS. We have employed droplet digital PCR (ddPCR) to detect rare L1 loci in mosaic human genomes. Our assay allows for the detection of L1 insertions as rare as one cell in every 10,000. ddPCR represents a robust method to be used alongside HTS techniques for detecting, validating and quantitating rare L1 insertion events in tumors and other tissues. The online version of this article (doi:10.1186/s13100-014-0030-4) contains supplementary material, which is available to authorized users.
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DOI：
10.1073/pnas.88.16.7276
发表时间：
1991-08-01
影响因子：
11.1
作者：
HOLLAND, PM;ABRAMSON, RD;GELFAND, DH
通讯作者：
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DOI：
10.1073/pnas.0831042100
发表时间：
2003-04-29
影响因子：
11.1
作者：
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Kazazian, HH