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首页|Circularization of genes and chromosome by CRISPR in human cells
10.1101/304493
来源：bioRxiv_logobioRxiv

Circularization of genes and chromosome by CRISPR in human cells

Circularization of genes and chromosome by CRISPR in human cells

Lin Lin 1Devitt M?ller Henrik 2Xu Xun 3Regenberg Birgitte 2Luo Yonglun 4Liu Xin 3Xiang Xi 5Petersen Trine Skov 1Zhang Xiuqing 6Jensen Uffe Birk 7Yang Luhan 8Church George M. 9Wang Jian 3Bolund Lars 10Huang Jinrong 11Yang Huanming 3Kjeldsen Eigil12

1. Department of Biomedicine, Aarhus University 2. Department of Biology, Faculty of Science, University of Copenhagen 3. BGI-Shenzhen||BGI-Qingdao||China National GeneBank, BGI-Shenzhen 4. Department of Biomedicine, Aarhus University||BGI-Shenzhen||BGI-Qingdao||China National GeneBank, BGI-Shenzhen||Lars Bolund Institute of Regenerative Medicine, BGI-Qingdao 5. BGI Education Center, University of Chinese Academy of Sciences||Department of Biomedicine, Aarhus University||BGI-Qingdao 6. BGI Education Center, University of Chinese Academy of Sciences||BGI-Shenzhen||China National GeneBank, BGI-Shenzhen 7. Department of Biomedicine, Aarhus University||Department of Clinical Medicine, Aarhus University 8. eGenesis, Inc. 9. eGenesis, Inc.||Department of Genetics, Harvard Medical School 10. Department of Biomedicine, Aarhus University||BGI-Shenzhen||BGI-Qingdao||Lars Bolund Institute of Regenerative Medicine, BGI-Qingdao 11. Department of Biology, Faculty of Science, University of Copenhagen||BGI-Shenzhen||BGI-Qingdao||China National GeneBank, BGI-Shenzhen 12. Department of Clinical Medicine, Aarhus University

遗传学分子生物学生物工程学

Lin Lin,Devitt M?ller Henrik,Xu Xun,Regenberg Birgitte,Luo Yonglun,Liu Xin,Xiang Xi,Petersen Trine Skov,Zhang Xiuqing,Jensen Uffe Birk,Yang Luhan,Church George M.,Wang Jian,Bolund Lars,Huang Jinrong,Yang Huanming,Kjeldsen Eigil.Circularization of genes and chromosome by CRISPR in human cells[EB/OL].(2025-03-28)[2025-09-24].https://www.biorxiv.org/content/10.1101/304493.点此复制

Abstract Extrachromosomal circular DNA (eccDNA) and ring chromosomes are genetic alterations found in humans with genetic disorders and diseases such as cancer. However, there is a lack of genetic engineering tool to recapitulate these features. Here, we report the discovery that delivery of pairs of CRISPR/Cas9 guide RNAs into human cells generate functional eccDNAs and ring chromosomes. We generated a dual-fluorescence eccDNA biosensor system, which allows us to study CRISPR deletion, inversion, and circularization of genes inside cells. Analysis after CRISPR editing at intergenic and genic loci in human embryonic kidney 293T cells and human mammary fibroblasts reveal that CRISPR deleted DNA readily form eccDNA in human cells. DNA in sizes from a few hundred base pairs up to a 47.4 megabase-sized ring chromosome (chr18) can be circularized. Our discoveries advance and expand CRISPR-Cas9 technology applications for genetic engineering, modeling of human diseases, and chromosome engineering. One Sentence Summary: CRISPR circularization of DNA offers new tools for studying eccDNA biogenesis, function, chromosome engineering, and synthetic biology.
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