CRISPR-C:circularization of genes and chromosome by CRISPR in human cells
| العنوان: | CRISPR-C:circularization of genes and chromosome by CRISPR in human cells |
|---|---|
| المؤلفون: | Xun Xu, Yonglun Luo, George M. Church, Eigil Kjeldsen, Henrik Møller, Xin Liu, Xiuqing Zhang, Birgitte Regenberg, Jian Wang, Luhan Yang, Jinrong Huang, Uffe Birk Jensen, Xi Xiang, Huanming Yang, Trine Skov Petersen, Lin Lin, Lars Bolund |
| المصدر: | Møller, H D, Lin, L, Xiang, X, Petersen, T S, Huang, J, Yang, L, Kjeldsen, E, Jensen, U B, Zhang, X, Liu, X, Xu, X, Wang, J, Yang, H, Church, G M, Bolund, L, Regenberg, B & Luo, Y 2018, ' CRISPR-C : circularization of genes and chromosome by CRISPR in human cells ', Nucleic Acids Research, vol. 46, no. 22, pp. e131 . https://doi.org/10.1093/nar/gky767Test Nucleic Acids Research Møller, H D, Lin, L, Xiang, X, Petersen, T S, Huang, J, Yang, L, Kjeldsen, E, Jensen, U B, Zhang, X, Liu, X, Xu, X, Wang, J, Yang, H, Church, G M, Bolund, L, Regenberg, B & Luo, Y 2018, ' CRISPR-C : circularization of genes and chromosome by CRISPR in human cells ', Nucleic Acids Research, vol. 46, no. 22, e131, pp. 1-13 . https://doi.org/10.1093/nar/gky767Test |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, DNA End-Joining Repair, Genetic Vectors, Green Fluorescent Proteins, Computational biology, Biosensing Techniques, Extrachromosomal circular DNA, Biology, Genome, Cell Line, 03 medical and health sciences, Genome editing, Genes, Reporter, CRISPR-Associated Protein 9, Genetics, CRISPR, Humans, Clustered Regularly Interspaced Short Palindromic Repeats, Guide RNA, Gene, Fluorescent Dyes, Gene Editing, 030102 biochemistry & molecular biology, Base Sequence, Cas9, Genome, Human, Chromosome, Fibroblasts, Luminescent Proteins, 030104 developmental biology, HEK293 Cells, Genetic Loci, Methods Online, CRISPR-Cas Systems, DNA, Circular, Chromosomes, Human, Pair 18, RNA, Guide, Kinetoplastida |
| الوصف: | 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. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::53e8ee99fdcad0d52909553f178d5b13Test https://pure.au.dk/portal/da/publications/crisprcTest(74a8ae42-2325-49a0-97a8-559bb7df8670).html |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....53e8ee99fdcad0d52909553f178d5b13 |
| قاعدة البيانات: | OpenAIRE |
| الوصف غير متاح. |