Cancer translocations in human cells induced by zinc finger and TALE nucleases.
Cancer translocations in human cells induced by zinc finger and TALE nucleases.
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DOI：
10.1101/gr.147314.112
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发表时间：
2013-07
期刊：
影响因子：
7
通讯作者：
Brunet E
中科院分区：
文献类型：
--
作者：
Piganeau M;Ghezraoui H;De Cian A;Guittat L;Tomishima M;Perrouault L;René O;Katibah GE;Zhang L;Holmes MC;Doyon Y;Concordet JP;Giovannangeli C;Jasin M;Brunet E
染色体易位是许多癌症的特征，并导致作为癌基因的融合基因的表达。然而，在癌症中发现的丰富的基因组畸变使得将特定表型变化分配给特定畸变具有挑战性。在这项研究中，我们开始使用锌指（ZFN）和转录激活因子样效应物（TALEN）核酸酶的基因组编辑来重新设计人类细胞中同源内源性基因座的易位相关癌基因。使用ZFN和TALEN设计在相关易位断点精确切割，我们分别诱导尤文肉瘤和间变性大细胞淋巴瘤（ALCL）中发现的癌症相关t（11;22）（q24;q12）和t（2;5）（p23;q35）易位。我们以高效率恢复了两种易位，导致EWSR 1-FLI 1和NPM 1-ALK融合体的表达。在人胚胎干（ES）细胞来源的间充质前体细胞中ZFN裂解后恢复的断点连接完全重现了尤文肉瘤患者肿瘤细胞中发现的基因组特征。这种使用定制核酸酶的方法表明，可以从天然启动子诱导癌细胞中发现的融合基因的表达，从而可以询问人类细胞中肿瘤相关易位的潜在机制和致癌后果。采用类似的策略，在患者细胞系中逆转ALCL易位，以恢复两条参与染色体的完整性，进一步扩大可通过定制核酸酶工程改造的基因组重排库。
Chromosomal translocations are signatures of numerous cancers and lead to expression of fusion genes that act as oncogenes. The wealth of genomic aberrations found in cancer, however, makes it challenging to assign a specific phenotypic change to a specific aberration. In this study, we set out to use genome editing with zinc finger (ZFN) and transcription activator-like effector (TALEN) nucleases to engineer, de novo, translocation-associated oncogenes at cognate endogenous loci in human cells. Using ZFNs and TALENs designed to cut precisely at relevant translocation breakpoints, we induced cancer-relevant t(11;22)(q24;q12) and t(2;5)(p23;q35) translocations found in Ewing sarcoma and anaplastic large cell lymphoma (ALCL), respectively. We recovered both translocations with high efficiency, resulting in the expression of the EWSR1–FLI1 and NPM1–ALK fusions. Breakpoint junctions recovered after ZFN cleavage in human embryonic stem (ES) cell–derived mesenchymal precursor cells fully recapitulated the genomic characteristics found in tumor cells from Ewing sarcoma patients. This approach with tailored nucleases demonstrates that expression of fusion genes found in cancer cells can be induced from the native promoter, allowing interrogation of both the underlying mechanisms and oncogenic consequences of tumor-related translocations in human cells. With an analogous strategy, the ALCL translocation was reverted in a patient cell line to restore the integrity of the two participating chromosomes, further expanding the repertoire of genomic rearrangements that can be engineered by tailored nucleases.
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影响因子：
15.8
作者：
Barberi T;Willis LM;Socci ND;Studer L
通讯作者：
Studer L
影响因子：
46.9
作者：
通讯作者：
--
DOI：
10.1073/pnas.0603507103
发表时间：
2006-06-27
影响因子：
11.1
作者：
Kasprzycka, Monika;Marzec, Michal;Wasik, Mariusz A.
通讯作者：
Wasik, Mariusz A.
影响因子：
2.8
作者：
Chansky, HA;Barahmand-pour, F;Yang, L
通讯作者：
Yang, L
影响因子：
7
作者：
Lee, Hyung Joo;Kim, Eunji;Kim, Jin-Soo
通讯作者：
Kim, Jin-Soo