Bacterial transfer of large functional genomic dna into human cells

ABSTRACT Efficient transfer of chromosome-based vectors into mammalian cells is difficult, mostly due to their large size. Using a genetically engineered invasive Escherichia coli vector,

alpha satellite DNA cloned in P1-based artificial chromosome was stably delivered into the HT1080 cell line and efficiently generated human artificial chromosomes de novo. Similarly, a large

genomic cystic fibrosis transmembrane conductance regulator (CFTR) construct of 160 kb containing a portion of the CFTR gene was stably propagated in the bacterial vector and transferred

into HT1080 cells where it was transcribed, and correctly spliced, indicating transfer of an intact and functional locus of at least 80 kb. These results demonstrate that bacteria allow the

cloning, propagation and transfer of large intact and functional genomic DNA fragments and their subsequent direct delivery into cells for functional analysis. Such an approach opens new

perspectives for gene therapy. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS OPTIONS Access through your

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FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS PANEL OF HUMAN CELL LINES WITH HUMAN/MOUSE ARTIFICIAL CHROMOSOMES Article Open access 22 February 2022 EFFICIENT

TARGETED TRANSGENESIS OF LARGE DONOR DNA INTO MULTIPLE MOUSE GENETIC BACKGROUNDS USING BACTERIOPHAGE BXB1 INTEGRASE Article Open access 31 March 2022 FIND AND CUT-AND-TRANSFER (FICAT)

MAMMALIAN GENOME ENGINEERING Article Open access 03 December 2021 ACCESSION CODES ACCESSIONS GENBANK/EMBL/DDBJ * accession number AY299332 * accession numbers AJ563631–AJ563650 * EMBL /

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Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS This work was supported in part by two grants from l'association Vaincre la Mucoviscidose in 2002–2003 and 2003–2004,

the latter together with the German Association Mukoviszidose e.v. D Schindelhauer was supported by the Deutsche Forschung Gemeinschaft and MD Amaral by research Grant POCTI / 1999 / MGI /

35737 from FCT Portugal. AS Ramalho was a recipient of PhD fellowship SFRH / BD / 3085 / 2000 from FCT, Portugal. We thank C Klein and M Speicher for FISH equipment. AUTHOR INFORMATION

Author notes * A Laner and S Goussard: These authors contributed equally to this work AUTHORS AND AFFILIATIONS * Department of Medical Genetics, Childrens Hospital, Ludwig Maximilians

University, Munich, Germany A Laner, T Schwarz & D Schindelhauer * Unité des Agents Antibactériens, Institut Pasteur, Paris, France S Goussard, P Courvalin & C Grillot-Courvalin *

Centre of Human Genetics, National Institute of Health Dr Ricardo Jorge, Lisboa, Portugal A S Ramalho & M D Amaral * Department of Chemistry and Biochemistry, Faculty of Sciences,

University of Lisboa, Lisboa, Portugal M D Amaral * Institute of Human Genetics, Technical University, Munich, Germany D Schindelhauer * Livestock Biotechnology, Life Sciences Center

Weihenstephan, Freising, Germany D Schindelhauer Authors * A Laner View author publications You can also search for this author inPubMed Google Scholar * S Goussard View author publications

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author publications You can also search for this author inPubMed Google Scholar * D Schindelhauer View author publications You can also search for this author inPubMed Google Scholar * C

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Laner, A., Goussard, S., Ramalho, A. _et al._ Bacterial transfer of large functional genomic DNA into human cells. _Gene Ther_ 12, 1559–1572 (2005). https://doi.org/10.1038/sj.gt.3302576

Download citation * Received: 10 February 2005 * Accepted: 22 May 2005 * Published: 23 June 2005 * Issue Date: November 2005 * DOI: https://doi.org/10.1038/sj.gt.3302576 SHARE THIS ARTICLE

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by the Springer Nature SharedIt content-sharing initiative KEYWORDS * gene delivery * bacterial E. coli vector * human artificial chromosome * CFTR