Molecular Biology Reports

, Volume 47, Issue 2, pp 1491–1498 | Cite as

Highly efficient CRISPR-targeting of the murine Hipp11 intergenic region supports inducible human transgene expression

  • Jill Browning
  • Michael Rooney
  • Emily Hams
  • Satoru Takahashi
  • Seiya Mizuno
  • Fumihiro Sugiyama
  • Padraic G. Fallon
  • Vincent P. KellyEmail author
Short Communication


Safe harbor loci allow predicable integration of a transgene into the genome without perturbing endogenous gene activity and for decades have been exploited in the mouse to investigate gene function, generate humanised models and create tissue specific reporter and Cre recombinase expressing lines. Herein, we show that the murine Hipp11 intergenic region can facilitate highly efficient integration of a large transgene—the human CD1A promoter and coding region—by means of CRISPR-Cas9 mediated homology directed repair. The data shows that the single copy human CD1A transgene is faithfully expressed in an inducible manner in homozygous animals in both macrophage and dendritic cells. Our results validate the Hipp11 intergenic region as being a highly amenable target site for functional transgene integration in mouse.


Intergenic site 2 Isg2 Hipp11 H11 Safe-harbor locus Humanized mice CRISPR-Cas9 CD1A 



J.B. is a Postgraduate Research Scholar funded by the Irish Research Council under the Government of Ireland Programme (GOIPG/2015/3729), M.R. was a Masters of Immunology Student at Trinity College Dublin. This work was supported by the National Children Research Centre. E.H. is a postdoctoral researcher funded by an SFI starting investigator research grant (15/SIRG/3473).

Author Contributions

VPK and PGF conceived and designed the study. JB, MR, EH, SM, and FS performed the experiments. JB, MR, EH, SM, FS, VPK and PGF interpreted the results. JB and VPK wrote the manuscript.

Compliance with ethical standards

Conflicts of interest

The authors declare no conflict of interest.

Supplementary material

11033_2019_5204_MOESM1_ESM.docx (18 kb)
Supplementary material 1 (DOCX 17 kb)
11033_2019_5204_MOESM2_ESM.pptx (1.2 mb)
Supplementary material 2 Supplementary Figure 1. LCs were isolated from ear skin of WT mice and homozygous mH11hCD1a mice, either untreated or administered Aldara cream (containing imiquimod) to the ear for 6 consecutive days. Cells were gated on LIVE/DEAD aqua -ve cells, CD45+ve and CD11c +ve cells and presented as huCD1a-PB vs CD207-PE (PPTX 1219 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.School of Biochemistry& ImmunologyTrinity Biomedical Sciences Institute, Trinity College DublinDublin 2Ireland
  2. 2.School of MedicineTrinity Biomedical Sciences Institute, Trinity College DublinDublin 2Ireland
  3. 3.1-1-1 Tennodai Laboratory Animal Resource CenterUniversity of TsukubaTsukubaJapan

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