title: Engineering of Systematic Elimination of a Targeted Chromosome in Human Cells.
abstract: Embryonic trisomy leads to abortion or congenital genetic disorders in humans. The most common autosomal chromosome abnormalities are trisomy of chromosomes 13, 18, and 21. Although alteration of gene dosage is thought to contribute to disorders caused by extra copies of chromosomes, genes associated with specific disease phenotypes remain unclear. To generate a normal cell from a trisomic cell as a means of etiological analysis or candidate therapy for trisomy syndromes, we developed a system to eliminate a targeted chromosome from human cells. Chromosome 21 was targeted by integration of a DNA cassette in HeLa cells that harbored three copies of chromosome 21. The DNA cassette included two inverted loxP sites and a herpes simplex virus thymidine kinase (HSV-tk) gene. This system causes missegregation of chromosome 21 after expression of Cre recombinase and subsequently enables the selection of cells lacking the chromosome by culturing in a medium that includes ganciclovir (GCV). Cells harboring only two copies of chromosome 21 were efficiently induced by transfection of a Cre expression vector, indicating that this approach is useful for eliminating a targeted chromosome.
authors.affiliation: Pediatric Oral Medicine, Division of Oral Health, Growth & Development, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
authors.forename: Hiroshi
authors.identifier: 0000-0002-6575-372X
authors.initials: H
authors.lastname: Sato
authors.affiliation: Pediatric Oral Medicine, Division of Oral Health, Growth & Development, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kato
authors.affiliation: Pediatric Oral Medicine, Division of Oral Health, Growth & Development, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
authors.forename: Haruyoshi
authors.identifier: 0000-0003-1138-3943
authors.initials: H
authors.lastname: Yamaza
authors.affiliation: Pediatric Dentistry & Special Needs Dentistry, Kyushu University Hospital, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
authors.forename: Keiji
authors.identifier: 0000-0002-4279-7930
authors.initials: K
authors.lastname: Masuda
authors.affiliation: Pediatric Oral Medicine, Division of Oral Health, Growth & Development, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
authors.forename: Huong Thi Nguyen
authors.initials: HT
authors.lastname: Nguyen
authors.affiliation: Pediatric Oral Medicine, Division of Oral Health, Growth & Development, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
authors.forename: Thanh Thi Mai
authors.initials: TT
authors.lastname: Pham
authors.affiliation: Pediatric Oral Medicine, Division of Oral Health, Growth & Development, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
authors.forename: Xu
authors.initials: X
authors.lastname: Han
authors.affiliation: Pediatric Oral Medicine, Division of Oral Health, Growth & Development, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Hirofuji
authors.affiliation: Pediatric Oral Medicine, Division of Oral Health, Growth & Development, Faculty of Dental Science, Kyushu University, Maidashi 3-1-1, Higashi-Ku, Fukuoka 812-8582, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Nonaka
mesh_terms: D002872:Chromosome Deletion; D025063:Chromosome Disorders / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D002882:Chromosomes, Human, Pair 13 / Q000235:genetics; D002887:Chromosomes, Human, Pair 18 / Q000235:genetics; D002891:Chromosomes, Human, Pair 21 / Q000235:genetics; D015774:Ganciclovir / Q000737:chemistry; D018628:Gene Dosage; D015316:Genetic Therapy; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D019426:Integrases / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection; D014314:Trisomy / Q000235:genetics* / Q000473:pathology
chemical_list: D013937:Thymidine Kinase; C045073:Cre recombinase; D019426:Integrases; D015774:Ganciclovir