CRISPR-C: circularization of genes and chromosome by CRISPR in human cells

Henrik Devitt Møller, Lin Lin, Xi Xiang, Trine Skov Petersen, Jinrong Huang, Luhan Yang, Eigil Kjeldsen, Uffe Birk Jensen, Xiuqing Zhang, Xin Liu, Xun Xu, Jian Wang, Huanming Yang, George M Church, Lars Bolund, Birgitte Regenberg, Yonglun Luo

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review


Extrachromosomal circular DNA (eccDNA) and ring chromosomes are genetic alterations found in humans with genetic disorders. However, there is a lack of genetic engineering tools to recapitulate and study the biogenesis of eccDNAs. Here, we created a dual-fluorescence biosensor cassette, which upon the delivery of pairs of CRISPR/Cas9 guide RNAs, CRISPR-C, allows us to study the biogenesis of a specific fluorophore expressing eccDNA in human cells. We show that CRISPR-C can generate functional eccDNA, using the novel eccDNA biosensor system. We further reveal that CRISPR-C also can generate eccDNAs from intergenic and genic loci in human embryonic kidney 293T cells and human mammary fibroblasts. EccDNAs mainly forms by end-joining mediated DNA-repair and we show that CRISPR-C is able to generate endogenous eccDNAs in sizes from a few hundred base pairs and ranging up to 207 kb. Even a 47.4 megabase-sized ring chromosome 18 can be created by CRISPR-C. Our study creates a new territory for CRISPR gene editing and highlights CRISPR-C as a useful tool for studying the cellular impact, persistence and function of eccDNAs.

Original languageEnglish
Article numbere131
JournalNucleic Acids Research
Pages (from-to)e131
Publication statusPublished - 14 Dec 2018


Dive into the research topics of 'CRISPR-C: circularization of genes and chromosome by CRISPR in human cells'. Together they form a unique fingerprint.

Cite this