title: Novel method to rescue a lethal phenotype through integration of target gene onto the X-chromosome.
abstract: The loss-of-function mutations of serine protease inhibitor, Kazal type 1 (SPINK1) gene are associated with human chronic pancreatitis, but the underlying mechanisms remain unknown. We previously reported that mice lacking Spink3, the murine homologue of human SPINK1, die perinatally due to massive pancreatic acinar cell death, precluding investigation of the effects of SPINK1 deficiency. To circumvent perinatal lethality, we have developed a novel method to integrate human SPINK1 gene on the X chromosome using Cre-loxP technology and thus generated transgenic mice termed "X-SPINK1". Consistent with the fact that one of the two X chromosomes is randomly inactivated, X-SPINK1 mice exhibit mosaic pattern of SPINK1 expression. Crossing of X-SPINK1 mice with Spink3+/-mice rescued perinatal lethality, but the resulting Spink3-/-;XXSPINK1mice developed spontaneous pancreatitis characterized by chronic inflammation and fibrosis. The results show that mice lacking a gene essential for cell survival can be rescued by expressing this gene on the X chromosome. The Spink3-/-;XXSPINK1mice, in which this method has been applied to partially restore SPINK1 function, present a novel genetic model of chronic pancreatitis.
authors.affiliation: Institute of Resource Development and Analysis, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.|Department of Gastroenterological Surgery, Kumamoto University, 1-1-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Sakata
authors.affiliation: Institute of Resource Development and Analysis, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.
authors.forename: Kimi
authors.initials: K
authors.lastname: Araki
authors.affiliation: Department of Biochemistry, Toho University School of Medicine, 5-21-16 Omori-Nishi, Ota-ku, Tokyo 143-8540, Japan.
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Nakano
authors.affiliation: Department of Biochemistry, Toho University School of Medicine, 5-21-16 Omori-Nishi, Ota-ku, Tokyo 143-8540, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Nishina
authors.affiliation: Department of Biochemistry, Toho University School of Medicine, 5-21-16 Omori-Nishi, Ota-ku, Tokyo 143-8540, Japan.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Komazawa-Sakon
authors.affiliation: Department of Biochemistry, Toho University School of Medicine, 5-21-16 Omori-Nishi, Ota-ku, Tokyo 143-8540, Japan.
authors.forename: Shin
authors.initials: S
authors.lastname: Murai
authors.affiliation: Department of Medicine, David Geffen School of Medicine at the University of California Los Angeles, and VA Greater Los Angeles Healthcare System, Los Angeles, CA, USA.
authors.forename: Grace E
authors.initials: GE
authors.lastname: Lee
authors.affiliation: Department of Gastroenterological Surgery, Kumamoto University, 1-1-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Hashimoto
authors.affiliation: Department of Cellular and Molecular Neuropathology, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-Ku, Tokyo 113-8421, Japan.
authors.forename: Chigure
authors.initials: C
authors.lastname: Suzuki
authors.affiliation: Department of Cellular and Molecular Neuropathology, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-Ku, Tokyo 113-8421, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Uchiyama
authors.affiliation: Department of Pathology, Kurashiki Central Hospital, 1-1-1 Miwa, Kurashiki, Okayama 710-8602, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Notohara
authors.affiliation: Department of Medicine, David Geffen School of Medicine at the University of California Los Angeles, and VA Greater Los Angeles Healthcare System, Los Angeles, CA, USA.
authors.forename: Anna S
authors.initials: AS
authors.lastname: Gukovskaya
authors.affiliation: Department of Medicine, David Geffen School of Medicine at the University of California Los Angeles, and VA Greater Los Angeles Healthcare System, Los Angeles, CA, USA.
authors.forename: Ilya
authors.initials: I
authors.lastname: Gukovsky
authors.affiliation: Institute of Resource Development and Analysis, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.
authors.forename: Ken-Ichi
authors.initials: KI
authors.lastname: Yamamura
authors.affiliation: Department of Gastroenterological Surgery, Kumamoto University, 1-1-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Baba
authors.affiliation: Institute of Resource Development and Analysis, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Ohmuraya
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D055879:Gene Knock-In Techniques; D006023:Glycoproteins / Q000172:deficiency*; D006801:Humans; D019426:Integrases; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D010195:Pancreatitis / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D029584:Prostatic Secretory Proteins; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D014960:X Chromosome / Q000235:genetics / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D029584:Prostatic Secretory Proteins; C486642:SPINK1 protein, human; C487134:Spink1 protein, mouse; D014359:Trypsin Inhibitor, Kazal Pancreatic; C045073:Cre recombinase; D019426:Integrases