title: Red/ET recombination with chimeric oligonucleotides allows rapid generation of BAC transgenes harboring full-length or truncated huntingtin cDNA.
abstract: Huntington's disease (HD) is a fatal neurodegenerative disorder that is caused by a CAG repeat expansion encoding a polyglutamine tract in the huntingtin (htt) gene. None of the existing HD mouse models recapitulate the exact disease symptoms and course as it is seen in humans and the generation of further HD disease models is challenging because of the size and complexity of the htt gene locus. Starting from a single substrate plasmid harboring human htt cDNA comprising 98 glutamine (Q) residues, we applied Red/ET recombination to generate four BDNF-BAC transgenes harboring full-length or truncated (N171) htt cDNA comprising 98 or 15 Q residues. BDNF (brain-derived neurotrophic factor) is expressed in the cortical neurons projecting to the striatal medium spiny neurons, and was used to direct htt transgene expression to investigate the contribution of these cell types to HD.
authors: Gene Bridges GmbH, Commercial Centre, Heidelberg, Germany. stefanie.hager@genebridges.com
authors: Stefanie
authors: S
authors: Hager
authors: Gene Bridges GmbH, Commercial Centre, Heidelberg, Germany.
authors: Saskia
authors: S
authors: Lösch
authors: Gene Bridges GmbH, Commercial Centre, Heidelberg, Germany.
authors: Stephan
authors: S
authors: Noll
authors: Mount Sinai Medical Center, Annenberg Building, New York, NY, USA.
authors: Loren
authors: L
authors: Khan-Vaughan
authors: Mount Sinai Medical Center, Annenberg Building, New York, NY, USA.
authors: Michelle E
authors: ME
authors: Ehrlich
authors: Gene Bridges GmbH, Commercial Centre, Heidelberg, Germany.
authors: Harald
authors: H
authors: Kranz
mesh_terms: D000818:Animals; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D000071058:Huntingtin Protein; D006816:Huntington Disease; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D009841:Oligonucleotides; D011995:Recombination, Genetic / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; C086055:HTT protein, human; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D009841:Oligonucleotides
keywords: BAC transgene; BDNF; Huntington's disease; Red/ET recombination; counterselection; huntingtin; recombineering