Transgenic Research

, Volume 27, Issue 6, pp 551–558 | Cite as

Mid-facial developmental defects caused by the widely used LacZ reporter gene when expressed in neural crest-derived cells

  • Xiaoxi Wei
  • Min Hu
  • Fei LiuEmail author
Brief Communication


Reporter genes play important roles in transgenic research. LacZ is a widely used reporter gene that encodes Escherichia coli β-galactosidase, an enzyme that is well known for its ability to hydrolyze X-gal into a blue product. It is unknown whether transgenic LacZ has any adverse effects. R26R reporter mice, containing a LacZ reporter gene, were generated to monitor the in vivo recombination activity of various transgenic Cre recombinase via X-gal staining. P0-Cre is expressed in neural crest-derived cells, which give rise to the majority of the craniofacial bones. Herein, we report that 12% of the R26R reporter mice harboring P0-Cre had unexpected mid-facial developmental defects manifested by the asymmetrical growth of some facial bones, thus resulting in tilted mid-facial structure, shorter skull length, and malocclusion. Histological examination showed a disorganization of the frontomaxillary suture, which may at least partly explain the morphological defect in affected transgenic mice. Our data calls for the consideration of the potential in vivo adverse effects caused by transgenic β-galactosidase.


R26R LacZ β-galactosidase Reporter gene P0-Cre Wnt1-Cre Transgenic mouse 



We thank Neil Thomas for help in editing the manuscript. We also thank Drs. Marco Giovannini, Andrew McMahon, and Phillipe Soriano for genetically modified mouse lines. This work was supported by NIH (AR062030 to FL) and NSFC (81470764 to MH). Micro-CT work was partly supported by the P30 Core Center award to the University of Michigan from NIAMS (AR 69620).

Compliance with ethical standards

Conflict of interest

The authors declares that they have no conflict of interest.


  1. Abe T, Fujimori T (2013) Reporter mouse lines for fluorescence imaging. Dev Growth Differ 55:390–405. CrossRefPubMedGoogle Scholar
  2. Ansari AM et al (2016) Cellular GFP toxicity and immunogenicity: potential Confounders in in vivo cell tracking experiments. Stem Cell Rev 12:553–559. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Araki K, Araki M, Miyazaki J, Vassalli P (1995) Site-specific recombination of a transgene in fertilized eggs by transient expression of Cre recombinase. Proc Natl Acad Sci U S A 92:160–164CrossRefGoogle Scholar
  4. Casola S (2010) Mouse models for miRNA expression: the ROSA26 locus. Methods Mol Biol 667:145–163. CrossRefPubMedGoogle Scholar
  5. Chen CM, Krohn J, Bhattacharya S, Davies B (2011) A comparison of exogenous promoter activity at the ROSA26 locus using a PhiiC31 integrase mediated cassette exchange approach in mouse ES cells. PLoS ONE 6:e23376. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Chen G et al (2017) Specific and spatial labeling of P0-Cre versus Wnt1-Cre in cranial neural crest in early mouse embryos. Genesis. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Danielian PS, Muccino D, Rowitch DH, Michael SK, McMahon AP (1998) Modification of gene activity in mouse embryos in utero by a tamoxifen-inducible form of Cre recombinase. Curr Biol 8:1323–1326CrossRefGoogle Scholar
  8. Detrait ER, Bowers WJ, Halterman MW, Giuliano RE, Bennice L, Federoff HJ, Richfield EK (2002) Reporter gene transfer induces apoptosis in primary cortical neurons. Mol Ther 5:723–730. CrossRefPubMedGoogle Scholar
  9. Fang F, Sun S, Wang L, Guan JL, Giovannini M, Zhu Y, Liu F (2015a) Neural crest-specific TSC1 deletion in mice leads to sclerotic craniofacial bone lesion. J Bone Miner Res 30:1195–1205. CrossRefPubMedPubMedCentralGoogle Scholar
  10. Fang F, Wei X, Hu M, Liu F (2015b) A mouse model of craniofacial bone lesion of tuberous sclerosis complex. Musculoskelet Regen. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Feltri ML, D’Antonio M, Previtali S, Fasolini M, Messing A, Wrabetz L (1999) P0-Cre transgenic mice for inactivation of adhesion molecules in Schwann cells. Ann N Y Acad Sci 883:116–123CrossRefGoogle Scholar
  12. Giovannini M et al (2000) Conditional biallelic Nf2 mutation in the mouse promotes manifestations of human neurofibromatosis type 2. Genes Dev 14:1617–1630PubMedPubMedCentralGoogle Scholar
  13. He Y, Sun X, Wang L, Mishina Y, Guan JL, Liu F (2017) Male germline recombination of a conditional allele by the widely used Dermo1-cre (Twist2-cre) transgene. Genesis. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Janbandhu VC, Moik D, Fassler R (2014) Cre recombinase induces DNA damage and tetraploidy in the absence of loxP sites. Cell Cycle 13:462–470. CrossRefPubMedGoogle Scholar
  15. Jiang X, Iseki S, Maxson RE, Sucov HM, Morriss-Kay GM (2002) Tissue origins and interactions in the mammalian skull vault. Dev Biol 241:106–116. CrossRefPubMedGoogle Scholar
  16. Lexow J, Poggioli T, Sarathchandra P, Santini MP, Rosenthal N (2013) Cardiac fibrosis in mice expressing an inducible myocardial-specific Cre driver. Dis Model Mech 6:1470–1476. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Liu F, Woitge HW, Braut A, Kronenberg MS, Lichtler AC, Mina M, Kream BE (2004) Expression and activity of osteoblast-targeted Cre recombinase transgenes in murine skeletal tissues. Int J Dev Biol 48:645–653. CrossRefPubMedGoogle Scholar
  18. Minoux M, Rijli FM (2010) Molecular mechanisms of cranial neural crest cell migration and patterning in craniofacial development. Development 137:2605–2621. CrossRefPubMedGoogle Scholar
  19. Morriss-Kay GM, Wilkie AO (2005) Growth of the normal skull vault and its alteration in craniosynostosis: insights from human genetics and experimental studies. J Anat 207:637–653. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Senarath-Yapa K, Chung MT, McArdle A, Wong VW, Quarto N, Longaker MT, Wan DC (2012) Craniosynostosis: molecular pathways and future pharmacologic therapy. Organogenesis 8:103–113. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Soriano P (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet 21:70–71. CrossRefPubMedGoogle Scholar
  22. Wang L, Mishina Y, Liu F (2015) Osterix-Cre transgene causes craniofacial bone development defect. Calcif Tissue Int 96:129–137. CrossRefPubMedGoogle Scholar
  23. Wei X, Thomas N, Hatch NE, Hu M, Liu F (2017) Postnatal craniofacial skeletal development of female C57BL/6NCrl mice. Front Physiol 8:697. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Yamauchi Y et al (1999) A novel transgenic technique that allows specific marking of the neural crest cell lineage in mice. Dev Biol 212:191–203. CrossRefPubMedGoogle Scholar
  25. Zhao H, Feng J, Ho TV, Grimes W, Urata M, Chai Y (2015) The suture provides a niche for mesenchymal stem cells of craniofacial bones. Nat Cell Biol 17:386–396. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  1. 1.Department of OrthodonticsJilin University School and Hospital of StomatologyChangchunChina
  2. 2.Department of Biologic and Materials Sciences and Division of ProsthodonticsUniversity of Michigan School of DentistryAnn ArborUSA

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