RAISING is a high-performance method for identifying random transgene integration sites.
RAISING is a high-performance method for identifying random transgene integration sites.
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DOI：
10.1038/s42003-022-03467-w
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发表时间：
2022-06-02
影响因子：
5.9
通讯作者：
中科院分区：
文献类型：
--
作者：
自然病毒感染和使用病毒载体的治疗干预都有很大的恶性转化风险。监测感染细胞的克隆扩增对于检测癌症是重要的。在这里，我们开发了一种通过检测转基因整合位点来跟踪克隆的新方法。raise（不受基因组DNA污染干扰的整合位点快速扩增）是一种灵敏、廉价的替代方法。它与Sanger测序结合我们的CLOVA （Clonality Value）软件的兼容性对于那些无法获得昂贵的高通量测序的人来说至关重要。我们分析了688名感染逆转录病毒HTLV-1的个体样本，这种病毒会导致成人t细胞白血病/淋巴瘤（ATL）。我们定义了一个克隆值，以100%的敏感性和94.8%的特异性识别ATL患者，我们的纵向分析也证明了ATL风险评估的有效性。未来的研究将证实我们的技术的广泛适用性，特别是在新兴的基因治疗领域。结合raise和CLOVA，提出了一种检测和监测病毒和病毒载体克隆整合的有效方法。
Both natural viral infections and therapeutic interventions using viral vectors pose significant risks of malignant transformation. Monitoring for clonal expansion of infected cells is important for detecting cancer. Here we developed a novel method of tracking clonality via the detection of transgene integration sites. RAISING (Rapid Amplification of Integration Sites without Interference by Genomic DNA contamination) is a sensitive, inexpensive alternative to established methods. Its compatibility with Sanger sequencing combined with our CLOVA (Clonality Value) software is critical for those without access to expensive high throughput sequencing. We analyzed samples from 688 individuals infected with the retrovirus HTLV-1, which causes adult T-cell leukemia/lymphoma (ATL) to model our method. We defined a clonality value identifying ATL patients with 100% sensitivity and 94.8% specificity, and our longitudinal analysis also demonstrates the usefulness of ATL risk assessment. Future studies will confirm the broad applicability of our technology, especially in the emerging gene therapy sector. Integrating RAISING and CLOVA, an effective method for detection and monitoring clonal integration of viruses and viral vectors is presented.
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影响因子：
12.3
作者：
Firouzi S;López Y;Suzuki Y;Nakai K;Sugano S;Yamochi T;Watanabe T
通讯作者：
Watanabe T
影响因子：
8.6
作者：
Akari, H;Nam, KH;Yoshikawa, Y
通讯作者：
Yoshikawa, Y
影响因子：
158.5
作者：
Hacein-Bey-Abina, S;Le Deist, F;Leiva, L
通讯作者：
Leiva, L
影响因子：
1.5
作者：
Demontis, Maria A.;Hilburn, Silva;Taylor, Graham P.
通讯作者：
Taylor, Graham P.
影响因子：
3.8
作者：
GRAHAM, FL;SMILEY, J;NAIRN, R
通讯作者：
NAIRN, R