Bornert Olivier, Nyström Alexander
Department of Dermatology, Faculty of Medicine, Medical Center, University of Freiburg, Freiburg, Germany.
Methods Mol Biol. 2019;1944:3-15. doi: 10.1007/978-1-4939-9095-5_1.
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.
互补DNA（cDNA）的大小和相对较高的鸟嘌呤-胞嘧啶（GC）含量对大型胶原蛋白的高效靶向工程而言是挑战。能够修饰胶原蛋白具有基础生物学和治疗学方面的意义，因为这将有助于开展研究，精确描述胶原蛋白与特定相互作用伙伴的相互作用，探究个体致病突变的后果，并评估精准医学方法的治疗适用性。以Ⅶ型胶原蛋白为例，我们在此将描述一种快速、简单地修饰编码大型胶原蛋白的cDNA的策略。该方法具有灵活性，可用于创建点突变、小的或大的缺失以及DNA插入。
