文章：

TALEN-介导的基因裁剪作为分析多发性骨髓瘤细胞获得性突变功能的工具

TALEN-mediated genetic tailoring as a tool to analyze the function of acquired mutations in multiple myeloma cells

原文发布日期：2014-05-09

DOI: 10.1038/bcj.2014.32

类型: Original Article

开放获取: 是

英文摘要：

Multiple myeloma (MM) is a clonal plasma cell malignancy that is initiated by a number of mutations and the process of disease progression is characterized by further acquisition of mutations. The identification and functional characterization of these myelomagenic mutations is necessary to better understand the underlying pathogenic mechanisms in this disease. Recent advancements in next-generation sequencing have made the identification of most of these mutations a reality. However, the functional characterization of these mutations has been hampered by the lack of proper and efficient tools to dissect these mutations. Here we explored the possible utility of transcription activator-like effector nuclease (TALEN) genome engineering technology to tailoring the genome of MM cells. To test this possibility, we targeted the HPRT1 gene and found that TALENs are a very robust and efficient genome-editing tool in MM cells. Using cotransfected green fluorescent protein as an enrichment marker, single-cell subclones with desirable TALEN modifications in the HPRT1 gene were obtained in as little as 3–4 weeks of time. We believe that TALENs will greatly facilitate the functional study of somatic mutations in MM as well as other cancers.

摘要翻译： 

多发性骨髓瘤（MM）是一种克隆性浆细胞恶性肿瘤，由多种突变引发，其疾病进展过程以持续获得新突变为特征。识别这些骨髓瘤致病性突变并进行功能表征，对于深入理解该疾病的潜在致病机制至关重要。随着新一代测序技术的最新进展，大部分此类突变的识别已成为现实。然而，由于缺乏合适有效的工具来解析这些突变，其功能表征研究一直进展缓慢。本研究探索了转录激活因子样效应物核酸酶（TALEN）基因组工程技术在多发性骨髓瘤细胞基因组定制中的应用潜力。为验证这一可能性，我们以HPRT1基因为靶点，发现TALEN技术在MM细胞中是一种非常强大且高效的基因组编辑工具。通过共转染绿色荧光蛋白作为富集标记，仅需3-4周即可获得HPRT1基因中含有所需TALEN修饰的单细胞亚克隆。我们相信TALEN技术将极大推动多发性骨髓瘤及其他癌症中体细胞突变的功能研究。

原文链接：

TALEN-mediated genetic tailoring as a tool to analyze the function of acquired mutations in multiple myeloma cells