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从 Colletes gigas 昆虫基因组中发现的一种自然活跃的转座子，可作为一种有前途的新型基因转移工具。

A Naturally Active Spy Transposon Discovered from the Insect Genome of Colletes gigas as a Promising Novel Gene Transfer Tool.

机构信息

College of Animal Science & Technology, Yangzhou University, Yangzhou, Jiangsu, 225009, China.

School of Basic Medical Sciences, Shenzhen University Medical School, Shenzhen University, Shenzhen, Guangdong, 518055, China.

出版信息

Adv Sci (Weinh). 2024 Aug;11(29):e2400969. doi: 10.1002/advs.202400969. Epub 2024 May 22.

DOI:10.1002/advs.202400969
PMID:38774947
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11304231/
Abstract

Novel active DNA transposons, such as Spy transposons from the PHIS superfamily, are identified through bioinformatics in this study. The native transposases cgSpy and cvSpy displayed transposition activities of approximately 85% and 35% compared to the hyperactive piggyBac transposase (hyPB). The cgSpy transposon showed unique characteristics, including a lack of overproduction inhibition and reduced efficiency for insertion sizes between 3.1 to 8.5 kb. Integration preferences of cgSpy are found in genes and regulatory regions, making it suitable for genetic manipulation. Evaluation in T-cell engineering demonstrated that cgSpy-mediated chimeric antigen receptor (CAR) modification is comparable to the PB system, indicating its potential utility in cell therapy. This study unveils the promising application of the active native transposase, Spy, from Colletes gigas, as a valuable tool for genetic engineering, particularly in T-cell manipulation.

摘要

本研究通过生物信息学方法鉴定了新型活性 DNA 转座子，如 PHIS 超家族的 Spy 转座子。与超活性 piggyBac 转座酶 (hyPB) 相比，天然转座酶 cgSpy 和 cvSpy 的转座活性分别约为 85%和 35%。cgSpy 转座子具有独特的特征，包括缺乏过度产生抑制和降低插入大小在 3.1 到 8.5kb 之间的效率。cgSpy 的整合偏好存在于基因和调控区域中，使其适合遗传操作。在 T 细胞工程中的评估表明，cgSpy 介导的嵌合抗原受体 (CAR) 修饰可与 PB 系统相媲美，表明其在细胞治疗中的潜在用途。这项研究揭示了来自 Colletes gigas 的活性天然转座酶 Spy 的有前途的应用，作为遗传工程，特别是 T 细胞操作的有价值的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/802c19e6db3c/ADVS-11-2400969-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/9c967e33ba10/ADVS-11-2400969-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/f011f03c2b1a/ADVS-11-2400969-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/983523718e0d/ADVS-11-2400969-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/a9cdfc8fe924/ADVS-11-2400969-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/bd79893763f0/ADVS-11-2400969-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/c28e40e4e6a3/ADVS-11-2400969-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/802c19e6db3c/ADVS-11-2400969-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/9c967e33ba10/ADVS-11-2400969-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/f011f03c2b1a/ADVS-11-2400969-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/983523718e0d/ADVS-11-2400969-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/a9cdfc8fe924/ADVS-11-2400969-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/bd79893763f0/ADVS-11-2400969-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/c28e40e4e6a3/ADVS-11-2400969-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f9d/11304231/802c19e6db3c/ADVS-11-2400969-g008.jpg

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本文引用的文献

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Nucleic Acids Res. 2023 Feb 28;51(4):1843-1858. doi: 10.1093/nar/gkad005.
2
Engineered Sleeping Beauty transposase redirects transposon integration away from genes.工程化睡眠美人转座酶将转座子整合重定向到基因之外。
Nucleic Acids Res. 2022 Mar 21;50(5):2807-2825. doi: 10.1093/nar/gkac092.
3
Transposon-Mediated CD19 Chimeric Antigen Receptor-T Cells Derived From CD45RA-Positive Peripheral Blood Mononuclear Cells Possess Potent and Sustained Antileukemic Function.
转座子介导的源自 CD45RA 阳性外周血单个核细胞的嵌合抗原受体 T 细胞具有强大且持久的抗白血病功能。
Front Immunol. 2022 Jan 27;13:770132. doi: 10.3389/fimmu.2022.770132. eCollection 2022.
4
Phase I clinical trial of EGFR-specific CAR-T cells generated by the piggyBac transposon system in advanced relapsed/refractory non-small cell lung cancer patients.猪内因子转座子系统生成的 EGFR 特异性 CAR-T 细胞治疗晚期复发/难治性非小细胞肺癌的 I 期临床试验。
J Cancer Res Clin Oncol. 2021 Dec;147(12):3725-3734. doi: 10.1007/s00432-021-03613-7. Epub 2021 May 25.
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CARAMBA: a first-in-human clinical trial with SLAMF7 CAR-T cells prepared by virus-free Sleeping Beauty gene transfer to treat multiple myeloma.卡拉巴：首例采用无病毒 Sleeping Beauty 基因转导制备的 SLAMF7 CAR-T 细胞治疗多发性骨髓瘤的人体临床试验。
Gene Ther. 2021 Sep;28(9):560-571. doi: 10.1038/s41434-021-00254-w. Epub 2021 Apr 13.
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Large granular lymphocytic leukemia - A retrospective study of 319 cases.大颗粒淋巴细胞白血病——319 例回顾性研究。
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