Related Experiment Video

Updated: Mar 12, 2026

Primordial Germ Cell Transplantation for CRISPR/Cas9-based Leapfrogging in Xenopus
05:34

Primordial Germ Cell Transplantation for CRISPR/Cas9-based Leapfrogging in Xenopus

Published on: February 1, 2018

8.8K

Novel method to rescue a lethal phenotype through integration of target gene onto the X-chromosome

Kazuya Sakata1,2, Kimi Araki1, Hiroyasu Nakano3

  • 1Institute of Resource Development and Analysis, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.

Scientific Reports
|November 16, 2016
PubMed

Insights

Loss-of-function mutations in the serine protease inhibitor, Kazal type 1 (SPINK1) gene are linked to chronic pancreatitis. A novel X-chromosome gene integration method in mice created a new genetic model for studying this disease.

Area of Science:

  • Genetics
  • Molecular Biology
  • Gastroenterology

Background:

  • Loss-of-function mutations in the serine protease inhibitor, Kazal type 1 (SPINK1) gene are associated with human chronic pancreatitis.
  • The precise mechanisms linking SPINK1 deficiency to pancreatitis remain unclear.
  • Murine Spink3 knockout mice exhibit perinatal lethality, hindering studies on SPINK1's role in pancreatic health.

Purpose of the Study:

  • To develop a novel method for studying SPINK1 deficiency in the pancreas.
  • To create a viable mouse model for investigating the role of SPINK1 in chronic pancreatitis.

Main Methods:

  • Developed a transgenic mouse model (X-SPINK1) by integrating human SPINK1 onto the X chromosome using Cre-loxP technology.
  • Generated Spink3 knockout mice with mosaic SPINK1 expression by crossing X-SPINK1 mice with Spink3+/- mice.
  • Observed the resulting Spink3-/-;XXSPINK1 mice for phenotypic changes.

Main Results:

  • The novel X-chromosome integration method successfully circumvented perinatal lethality associated with Spink3 deficiency.
  • Spink3-/-;XXSPINK1 mice exhibited mosaic SPINK1 expression due to random X chromosome inactivation.
  • These mice developed spontaneous chronic pancreatitis, characterized by inflammation and fibrosis, serving as a model for the disease.

Conclusions:

  • Expressing an essential gene on the X chromosome can rescue lethal genetic deficiencies.
  • The developed Spink3-/-;XXSPINK1 mouse model provides a valuable tool for understanding SPINK1's role in chronic pancreatitis pathogenesis.
  • This study establishes a novel genetic approach for studying essential genes with potential applications in disease modeling.

More Related Videos

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation09:48

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation

Published on: August 21, 2010

12.5K
Pooled shRNA Screen for Reactivation of MeCP2 on the Inactive X Chromosome11:15

Pooled shRNA Screen for Reactivation of MeCP2 on the Inactive X Chromosome

Published on: March 2, 2018

7.7K

Related Concept Videos

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
17.5K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
7.2K
Dosage Compensation02:50

Dosage Compensation

In animals, gender is determined by the number and type of sex chromosome. For example, human females have two X chromosomes, and males have one X and one Y chromosome, whereas C.elegans with one X chromosome is a male, and the one with two X chromosomes is a hermaphrodite.
In addition to sexual development, the X chromosome has genes involved in autosomal functions such as brain development and the immune system. Therefore, males and females with  distinct numbers of X chromosomes will...
7.7K
X-Inactivation01:58

X-Inactivation

The human X chromosome contains over ten times the number of genes as in the Y chromosome. Since males have only one X chromosome, and females have two, one might expect females to produce twice as many of the proteins, with undesirable results.
42.9K
The Ratio of X Chromosome to Autosomes02:45

The Ratio of X Chromosome to Autosomes

In most organisms, sex is determined by the ratio of X and Y chromosomes. However, in some organisms, such as Drosophila and C.elegans, sex is determined by the ratio of the number of X chromosomes to the number of sets of autosomes. The Y chromosome in Drosophila is active but does not determine sex. It contains genes responsible for the production of sperms in adult flies.  
Normal male Drosophila has a ratio of one X chromosome to two sets of autosomes. In contrast, normal female...
10.0K
Lethal Alleles02:41

Lethal Alleles

Agouti: A Lethal Allele
Lucien Cuénot discovered lethal alleles in 1905 while studying the inheritance of coat color in mice. The agouti gene is responsible for the color of the coat in mice. This gene codes for an agouti-signaling protein, which is responsible for melanin distribution in mammals. The wild-type allele gives rise to gray-brown coat color in mice, while the mutant allele gives rise to yellow coat color. In addition to coat color, the agouti gene is associated with the yellow...
18.8K