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Cloning and Mutagenesis Strategies for Large Collagens

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Collagen

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1944))

Abstract

The size and relatively high GC content of cDNAs are challenges for efficient targeted engineering of large collagens. There are both basic biological and therapeutic interests in the ability to modify collagens, as this would allow for studies precisely describing interactions of collagens with specific interaction partners, addressing consequences of individual disease-causing mutations, and assessing therapeutic applicability of precision medicine approaches. Using collagen VII as an example, we will here describe a strategy for rapid and simple modification of cDNAs encoding large collagens. The method is flexible and can be used for the creation of point mutations, small or large deletions, and insertion of DNA.

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Acknowledgments

Part of the work was financed by a research collaboration with ProQR, Netherlands. AN’s research was supported by grants from the German Research Foundation, DFG (grants NY90/2-1, NY90/3-2, SFB850-B11 to AN, BR1475/12-1 to LBT), and from the Dystrophic Epidermolysis Bullosa Research Association (DEBRA) (grant Nystrom Bruckner-Tuderman 1) to AN.

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Authors and Affiliations

  1. Department of Dermatology, Faculty of Medicine, Medical Center, University of Freiburg, Freiburg, Germany

    Olivier Bornert & Alexander Nyström

Authors
  1. Olivier Bornert
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  2. Alexander Nyström
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Corresponding author

Correspondence to Alexander Nyström .

Editor information

Editors and Affiliations

  1. Department of Biological Regulation, Weizmann Institute of Science, Rehovot, Israel

    Irit Sagi

  2. Department of Biological Regulation, Weizmann Institute of Science, Rehovot, Israel

    Nikolaos A. Afratis

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Bornert, O., Nyström, A. (2019). Cloning and Mutagenesis Strategies for Large Collagens. In: Sagi, I., Afratis, N. (eds) Collagen. Methods in Molecular Biology, vol 1944. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9095-5_1

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  • DOI: https://doi.org/10.1007/978-1-4939-9095-5_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9094-8

  • Online ISBN: 978-1-4939-9095-5

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