System
The Exchangeable Gene Trap System Trap Vector Exchangeable System ES Cell Lines Isolation of Trap Clones Registration in EGTC Establishment of Trap Mouse Lines Deposition of Trap Lines in CARD Expression Analysis by X-gal Staining Reference
Description
X-gal staining was performed for detecting β-galactocidase reporter expression in a wide variety of adult tissues of each trap line.
X-gal Staining Procedure
Materials Fixative Solution
2.7% formalin
0.2% glutaraldehyde
0.02% Nonidet P-40
in PBS
X-gal Stock Solution
2% X-gal (5-Bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) in
dimethylformamide
Store at -20℃ in small aliquots.
X-gal Working Solution
5mM Potassium ferricyanide
5mM Potassium ferrocyanide
0.5mM MgCl2
0.05% X-gal stock solution
in PBS
Make fresh just before use.
Procedure
- Place dissected adult tissues in PBS and cut them with a blade.
- Transfer tissues in small dishes or multi-well plates.
- Fix with the fixative solution for 30 minutes.
- Rinse tissues with 1%Triton X-100/PBS 10minutes.
- Rinse tissues with 2 changes of PBS for 10minutes each.
- Incubate tissues in X-gal working solution overnight.
- Post-fix with the fixative solution for 30 minutes.
- Rinse with 2 changes of PBS for 10minutes each.
Note: Tissues were rocked gently at room temperature on each step.
"NSrp70 is a novel nuclear speckle-related protein that
modulates alternative pre-mRNA splicing in vivo."
Kim, Y. D., Lee, J. Y., Oh, K. M., Araki, M., Araki, K., Yamamura, K.,
Jun, C. D.
Nucl. Acids Res., 39, 4300-4314 (2011). PubMed ID: 21296756.
"Comparative analysis of right element mutant lox sites on
recombination efficiency in embryonic stem cells." Araki, K.,
Okada, Y., Araki, M., Yamamura, K.
BMC Biotechnol., 10, 29 (2010). PubMed ID: 20356367.
"Defective development of the gall bladder and cystic duct
in Lgr4- hypomorphic mice."
Yamashita, R., Takegawa, Y., Sakumoto, M., Nakahara, M., Kawazu, H.,
Hoshii, T., Araki, K., Yokouchi, Y., Yamamura, K.
Dev. Dyn., 238, 993-1000, (2009). PubMed ID: 19301403.
"Basonuclin 2 has a function in the multiplication of
embryonic craniofacial mesenchymal cells and is orthologous to disco
proteins."
Vanhoutteghem, A., Maciejewski-Duval, A., Bouche, C., Delhomme, B.,
Herve, F., Daubigney, F., Soubigou, G., Araki, M., Araki, K., Yamamura,
K., Djian, P.
Proc. Natl. Acad. Sci. USA, 106, 14432-14437 (2009). PMID: 19706529.
"International Gene Trap Project: towards gene-driven
saturation mutagenesis in mice."
Araki, M., Araki, K., Yamamura, K.
Curr. Pharm. Biotechnol., 10, 221-229 (2009). PMID: 19199955.
"Early pre-implantation lethality in mice carrying truncated
mutation in the RNA polymerase 1-2 gene."
Chen, H., Li, Z., Haruna, K., Semba, K., Araki, M., Yamamura, K., Araki,
K.
Biochem. Biophys. Res. Commun., 365, 634-642 (2008). PMID: 18023416.
PMID: 18023416.
"Establishment of germline-competent embryonic stem cell
lines from the MSM/Ms strain."
Araki, K., Takeda, N., Yoshiki, A., Obata, Y., Nakagata, N., Shiroishi,
T., Moriwaki, K., Yamamura, K.
Mamm Genome, 20, 14-20 (2008). PubMed ID: 19082856.
"Gene trap mutagenesis in mice: new perspectives and tools
in cancer research."
Yamamura, K., Araki, K.
Cancer Sci.,99, 1-6 (2007). PubMed ID: 17877761.
"LGR4 regulates the postnatal development and integrity of
male reproductive tracts in mice."
Hoshii, T., Takeo, T., Nakagata, N., Takeya, M., Araki, K., Yamamura, K.
Biol. Reprod., 76, 303-313 (2007). PubMed ID: 17079737.
"Negative selection with the Diphtheria toxin A fragment
gene improves frequency of Cre-mediated cassette exchange in ES
cells."
Araki, K., Araki, M., Yamamura, K.
J. Biochem., 140, 793-798 (2006). PMID: 17043056.
"Impaired expression of importin/karyopherin beta1 leads to
post-implantation lethality."
Miura, K., Yoshinobu, K., Imaizumi, T., Haruna, K., Miyamoto, Y.,
Yoneda, Y., Nakagata, N., Araki, M., Miyakawa, T., Yamamura, K., Araki,
K.
Biochem. Biophys. Res. Commun., 341, 132-138 (2006). PMID: 16414015.
"A novel murine gene, Sickle tail (Skt), linked to the
Danforth's short tail (Sd) locus, is required for normal development of
the intervertebral disc."
Semba, K., Araki, K., Li, Z., Matsumoto, K. I., Suzuki, M., Nakagata,
N., Takagi, K., Takeya, M., Yoshinobu, K., Araki, M., Imai, K., Abe, K.,
Yamamura, K.
Genetics, 172, 445-456 (2006). PMID: 16204209.
"Characterization of an exchangeable gene trap using pU-17
carrying a stop codon-betageo cassette."
Taniwaki, T., Haruna, K., Nakamura, H., Sekimoto, T., Oike, Y.,
Imaizumi, T., Saito, F., Muta, M., Soejima, Y., Utoh, A., Nakagata, N.,
Araki, M., Yamamura, K., Araki, K.
Dev. Growth Differ., 47 (3), 163-172 (2005). PMID: 15840001.
"Increase of smooth muscle cell migration and of intimal
hyperplasia in mice lacking the alpha/beta hydrolase domain containing 2
gene."
Miyata, K., Oike, Y., Hoshii, T., Maekawa, H., Ogawa, H., Suda, T.,
Araki, K., Yamamura, K.
Biochem. Biophys. Res. Commun., 329 (1), 296-304 (2005). PMID: 15721306.
"Serine-arginine-rich nuclear protein Luc7l regulates
myogenesis in mice."
Kimura, E., Hidaka, K., Kida, Y., Morisaki, H., Shirai, M., Araki, K.,
Suzuki, M., Yamamura, K. I., Morisaki, T.
Gene, 341, 41-47 (2004). PMID: 15474286.
"Generation and genetic modification of dendritic cells
derived from mouse embryonic stem cells."
Senju, S., Hirata, S., Matsuyoshi, H., Masuda, M., Uemura, Y., Araki,
K., Yamamura, K., Nishimura, Y.
Blood, 101 (9), 3501-3508 (2003). PMID: 12406878.
"Site-directed integration of the cre gene mediated by Cre
recombinase using a combination of mutant lox sites."
Araki, K., Araki, M., Yamamura, K.
Nuc. Acids Res., 30 (19), e103 (2002). PMID: 12364620.
"Mutant mice lacking Crk-II caused by the gene trap
insertional mutagenesis: Crk-II is not essential for embryonic
development."
Imaizumi, T., Araki, K., M., Miura, K., Araki, M., Suzuki, M., Terasaki,
H., Yamamura, K.
Biochem. Biophys. Res. Commun., 266, 569-574 (1999). PMID: 10600543.
"Exchangeable gene trap using the Cre/mutated lox
system."
Araki, K., Imaizumi, T., Sekimoto, T., Yoshinobu, K., Yoshimuta, J.,
Akizuki, M., Miura, K., Araki, M., Yamamura, K.
Cellular and Molecular Biology, 45, 737-750 (1999). PMID: 10512203.
"Mice homozygous for a truncated form of CREB-binding
protein exhibit defects in hematopoiesis and vasculo-angiogenesis."
Oike, Y., Takakura, N., Hata, A., Kaname, T., Akizuki, M., Yamaguchi,
Y., Yasue, H., Araki, K., Yamamura, K., Suda, T.
Blood, 93, 2771-2779 (1999). PMID: 10216070.
"Comparison of ES cell fate in sandwiched aggregates and
co-cultured aggregates during blastocyst formation by monitored GFP
expression."
Shimada, H., Kaname, T., Suzuki, M., Hitoshi, Y., Araki, K., Imaizumi,
T., Yamamura, K.
Mol. Reprod. Dev., 52, 376-382 (1999). PMID: 10092117.
"Truncated CBP protein leads to classical Rubinstein-Taybi
syndrome phenotypes in mice: implications for a dominant-negative
mechanism."
Oike, Y., Hata, A., Mamiya, T., Kaname, T., Noda, Y., Suzuki, M., Yasue,
H., Nabeshima, T., Araki, K.,Yamamura, K.
Hum. Mol. Genet., 8, 387-396 (1999). PMID: 9949198.
"Targeted integration of DNA using mutant lox sites in
embryonic stem cells."
Araki, K., Araki, M. and Yamamura, K.
Nuc. Acids Res., 25 (4), 868-872 (1997). PMID: 9016639.
"Efficiency of recombination by Cre transient expression in
embryonic stem cells: comparison of various promoters."
Araki, K., Imaizumi, T., Okuyama, K., Oike, Y., Yamamura, K.
J. Biochem., 122, 977-82 (1997). PMID: 9443813.
"Site-specific recombination of transgene in fertilized eggs
by transiently expressed Cre recombinase."
Araki, K., Araki, M., Miyazaki, J., and Vassalli, P.
Proc. Natl. Acad. Sci., 92, 160-164 (1995). PMID: 7816809.
"An efficient gene-trap method using poly A trap vectors and
characterization of gene-trap events."
Niwa, H., Araki, K., Kimura, S., Taniguchi, S., Wakasugi, S., Yamamura,
K.
J. Biochem., 113, 343-349 (1993). PMID: 8387481