Sensitive detection of Cre-mediated recombination using droplet digital PCR reveals Tg(BGLAP-Cre) and Tg(DMP1-Cre) are active in multiple non-skeletal tissues

Dasgupta, Krishnakali; Lessard, Samantha G.; Hann, Steven; Fowler, Megan; Robling, Alexander G.; Warman, Matthew L.

doi:10.1016/j.bone.2020.115674

Cited by 11 publications

(13 citation statements)

References 38 publications

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“…(161)(162)(163)167,244) Osteoblastic activity was lower in the 8kb-Dmp1-Cre line than in the 10kb Dmp1-Cre line. (244) The 10kb-Dmp1-Cre line also shows Cre activity in muscle, brain, kidney, (141,163,244) gut, (161) and bone marrow cells, (244) but not bone marrow adipocytes. (161) The 8kb-Dmp1-Cre line shows activity in the brain, but not in muscle, kidney, or heart.…”

Section: Cre Mice Targeting Osteocytesmentioning

confidence: 99%

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Sites of Cre-recombinase activity in mouse lines targeting skeletal cells

Couasnay

Madel

Lim

et al. 2020

Journal of Bone and Mineral Research

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The Cre/Lox system is a powerful tool in the biologist's toolbox, allowing loss-of-function and gain-of-function studies, as well as lineage tracing, through gene recombination in a tissue-specific and inducible manner. Evidence indicates, however, that Cre transgenic lines have a far more nuanced and broader pattern of Cre activity than initially thought, exhibiting "off-target" activity in tissues/cells other than the ones they were originally designed to target. With the goal of facilitating the comparison and selection of optimal Cre lines to be used for the study of gene function, we have summarized in a single manuscript the major sites and timing of Cre activity of the main Cre lines available to target bone mesenchymal stem cells, chondrocytes, osteoblasts, osteocytes, tenocytes, and osteoclasts, along with their reported sites of "off-target" Cre activity. We also discuss characteristics, advantages, and limitations of these Cre lines for users to avoid common risks related to overinterpretation or misinterpretation based on the assumption of strict cell-type specificity or unaccounted effect of the Cre transgene or Cre inducers.

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Section: Cre Mice Targeting Osteocytesmentioning

confidence: 99%

“…Tables 1–6 list the Cre lines discussed and include references of documents describing Cre activity in bone and off‐target sites, and original articles reporting the lines (knock‐in–based Cre lines in these tables are labeled with an asterisk). ( 22‐213 )…”

Section: Introductionmentioning

confidence: 99%

Sites of Cre-recombinase activity in mouse lines targeting skeletal cells

Couasnay

Madel

Lim

et al. 2020

Journal of Bone and Mineral Research

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“…35 A recent study showed that BglapCre also targeted a range of non-skeletal cell types in mice; hence, caution should be used when interpreting data obtained using these mice. 56 The most commonly used transgenic mouse model for relatively mature osteoblasts is driven by the mouse 2.3 kb promoter for col- 35 Recent scRNA-seq studies on BM obtained from Col2.3:tdTomato mice have revealed 3 distinct populations of cells that are targeted by Col2.3Cre. 36 These populations expressed detectable levels of Cxcl12 and Kitl; however, the expression levels of these two mRNA transcripts were much lower than those detected in the Lepr:tdTomato BM cells.…”

Section: Osteoblastsmentioning

confidence: 99%

“…There are a number of transgenic mouse strains that have been used to more specifically target mature osteoblasts, including osteocalcin ( Bglap ) 55 and a variety of mice that were generated using promoters that target type 1 collagen 35 . A recent study showed that Bglap Cre also targeted a range of non‐skeletal cell types in mice; hence, caution should be used when interpreting data obtained using these mice 56 . The most commonly used transgenic mouse model for relatively mature osteoblasts is driven by the mouse 2.3 kb promoter for collagen 1α1 (2.3 kb) [ Col1a1 (2.3kb), commonly referred to as Col2.3 ] 35 .…”

Section: Bone Marrow Microenvironment Cells and Transgenic Mice Used To Target These Cells In Vivomentioning

confidence: 99%

“…There are two different Dmp1 Cre transgenic lines, and both have been shown to target some other cell types, including a subset of CXCL12‐abundant reticular (CAR) cells, osteoblasts, and bone lining cells 35,57,58 . Furthermore, a recent study showed that some cell types in brain, muscle, and arteries were also targeted by Dmp1 Cre 56 . Any conclusions made from studies using Dmp1 Cre mice are therefore not restricted to the effects of osteocytes.…”

Section: Bone Marrow Microenvironment Cells and Transgenic Mice Used To Target These Cells In Vivomentioning

confidence: 99%

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Regulation of murine B lymphopoiesis by stromal cells

Kwang

Tjin

Purton

2021

Immunological Reviews

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During the past two decades, there has been a significant increase in our understanding of how B lymphopoiesis is extrinsically regulated by stromal cells. The information on the stromal cell types that are now known to have important roles in regulating B lymphopoiesis has largely been obtained from studies in mice. Many of these studies (such as specifically knocking out a gene of interest in a particular stromal cell type) cannot be done in humans. Hence, this review focuses on what we have learned about the extrinsic regulation of B lymphopoiesis from studies in mice.It should be noted that there are some differences in the regulation of B lymphocyte production between mice and humans, which have been extensively reviewed by Ichii et al 1 A major difference between the two species is the role of the spleen in B lymphopoiesis. As is discussed further below, the spleen is a major site for the maturation of B lymphocytes in mice; however, in humans, it is a crucial reservoir of memory B cells 2 but does not contribute to B lymphocyte maturation. Cell surface markers that identify each of the developing B lymphocyte populations are also different between mice and humans. 1 Nevertheless, the information gained from studies in mice is very valuable and, with

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Visualizing Cathepsin K‐Cre Expression at the Single‐Cell Level with GFP Reporters

et al. 2022

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The Cre/lox system is a fundamental tool for functional genomic studies, and a number of Cre lines have been generated to target genes of interest spatially and temporally in defined cells or tissues; this approach has greatly expanded our knowledge of gene functions. However, the limitations of this system have recently been recognized, and we must address the challenge of so‐called nonspecific/off‐target effects when a Cre line is utilized to investigate a gene of interest. For example, cathepsin K (Ctsk) has been used as a specific osteoclast marker, and Cre driven by its promoter is widely utilized for osteoclast investigations. However, Ctsk‐Cre expression has recently been identified in other cell types, such as osteocytes, periosteal stem cells, and tenocytes. To better understand Ctsk‐Cre expression and ensure appropriate use of this Cre line, we performed a comprehensive analysis of Ctsk‐Cre expression at the single‐cell level in major organs and tissues using two green fluorescent protein (GFP) reporters (ROSA nT‐nG and ROSA tdT) and a tissue clearing technique in young and aging mice. The expression profile was further verified by immunofluorescence staining and droplet digital RT‐PCR. The results demonstrate that Ctsk‐Cre is expressed not only in osteoclasts but also at various levels in osteoblast lineage cells and other major organs/tissues, particularly in the brain, kidney, pancreas, and blood vessels. Furthermore, Ctsk‐Cre expression increases markedly in the bone marrow, skeletal muscle, and intervertebral discs in aging mice. These data will be valuable for accurately interpreting data obtained from in vivo studies using Ctsk‐Cre mice to avoid potentially misleading conclusions. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

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Sensitive detection of Cre-mediated recombination using droplet digital PCR reveals Tg(BGLAP-Cre) and Tg(DMP1-Cre) are active in multiple non-skeletal tissues

Cited by 11 publications

References 38 publications

Sites of Cre-recombinase activity in mouse lines targeting skeletal cells

Sites of Cre-recombinase activity in mouse lines targeting skeletal cells

Regulation of murine B lymphopoiesis by stromal cells

Visualizing Cathepsin K‐Cre Expression at the Single‐Cell Level with GFP Reporters

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