Abstract
A novel protein LUZP with 3 leucine zipper motifs at its amino terminus is predominantly expressed in the adult brain. A modified gene targeting approach was employed in an attempt to establish in vitro and in vivo models in whichLuzp is knock-out (KO) for phenotype assessment and a reporter genelacZ is knock-in (KI) for tracing its expression. We report in this study the molecular cloning of theLuzp gene, its targeting vector construction andLuzp-KO/lacZ-KI embryonic stem (ES) clone selection. Since LUZP is also expressed in ES cells, the possibility of embryonic lethality cannot be excluded when attempting to establishLuzp-null mutant mice. We have therefore examined the development of homozygousLuzp-KO/lacZ-KI clones in nude mice. Tissue types derived from all three embryonic germ layers were observed in teratomas developed in nude mice. In situ X-gal staining further revealed restricted expression of LUZP in neural lineage cells.
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Bourtchuladze R, Frenguelli B, Blendy J, Cioffi D, Schutz G, Silva AJ. Deficient long-term memory in mice with a targeted mutation of the cAMP-responsive element-binding protein. Cell 79:59–68;1994.
Dobrzanski P, Ryseck RP, Bravo R. Both N-and C-terminal domains of RelB are required for full transactivation: Role of the N-terminal leucine zipper-like motif. Mol Cell Biol 13:1572–1582;1993.
Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13;1983.
Friedman AD. GADD153/CHOP, a DNA damage-inducible protein, reduced CAAT/enhancer binding protein activities and increased apoptosis in 32D c13 myeloid cells. Cancer Res 56:3250–3256;1996.
Hu RQ, Koh S, Torgerson T, Cole AJ. Neuronal stress and injury in C57/BL mice after systemic kainic acid administration. Brain Res 810:229–240;1998.
Hurst HC. Leucine Zippers: Transcription Factors, ed 3. London, Academic Press, 1996.
Imaki J, Yoshida K, Yamashita K. A developmental study of cyclic AMP-response element binding protein (CREB) by in situ hybridization histochemistry and immunocytochemistry in the rat neocortex. Brain Res 651:269–274;1994.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277:680–685;1970.
Lim K, Chae CB. A simple assay for DNA transfection by incubation of the cells in culture dishes with substrates for beta-galactosidase. Biotechniques 7:576–579;1989.
Mortensen RM, Conner DA, Chao S, Geisterfer-Lowrance AA, Seidman JG. Production of homozygous mutant ES cells with a single targeting construct. Mol Cell Biol 12:2391–2395;1992.
Reubinoff BE, Pera MF, Fong CY, Trounson A, Bongso A. Embryonic stem cell lines from human blastocysts: Somatic differentiation in vitro. Nat Biotechnol 18:399–404;2000.
Ron D, Habener JF. CHOP, a novel developmentally regulated nuclear protein that dimerizes with transcription factors C/EBP and LAP and functions as a dominant-negative inhibitor of gene transcription. Genes Dev 6:439–453;1992.
Sambrook J, Fritsch EF, Maniatis T. Plasmid vectors: Extraction and purification of plasmid DNA; in Molecular Cloning: A Laboratory Manual, ed 2. Cold Spring Harbor, Cold Spring Harbor Laboratory Press, 21–52;1989.
Sanger F, Nicklen S, Coulson A. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467;1977.
Shaywitz AJ, Greenberg ME. CREB: A stimulus-induced transcription factor activated by a diverse array of extracellular signals. Annu Rev Biochem 68:821–861;1999.
Shu SY, Ju G, Fan LZ. The glucose oxidase-DAB-nickel method in peroxidase histochemistry of the nervous system. Neurosci Lett 85:169–171;1988.
Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG. The unlabeled antibody enzyme method of immunohistochemistry: Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333;1970.
Sun DS, Chang AC, Jenkins NA, Gilbert DJ, Copeland NG, Chang NC. Identification, molecular characterization, and chromosomal localization of the cDNA encoding a novel leucine zipper motif-containing protein. Genomics 36:54–62;1996.
Tamura TA, Konishi Y, Makino Y, Mikoshiba K. Mechanisms of transcriptional regulation and neural gene expression. Neurochem Int 29:573–581;1996.
Thomson JA, Kalishman J, Golos TG, Durning M, Harris CP, Becker RA, Hearn JP. Isolation of a primate embryonic stem cell line. Proc Natl Acad Sci USA 92:7844–7848;1995.
Wu GY, Deisseroth K, Tsien RW. Activity-dependent CREB phosphorylation: Convergence of a fast, sensitive calmodulin kinase pathway and a slow, less sensitive mitogen-activated protein kinase pathway. Proc Natl Acad Sci USA 98:2808–2813;2001.
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Lee, M.WY., Chang, A.C., Sun, DS. et al. Restricted expression of LUZP in neural lineage cells: A study in embryonic stem cells. J Biomed Sci 8, 504–511 (2001). https://doi.org/10.1007/BF02256613
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DOI: https://doi.org/10.1007/BF02256613