BAC constructs in transgenic reporter mouse lines control efficient and specific LacZ expression in hypertrophic chondrocytes under the complete Col10a1 promoter

Sonja Gebhard, Takako Hattori, Eva Bauer, Michael R. Bösl, Britta Schlund, Ernst Pöschl, Nadia Adam, Benoit De Crombrugghe, Klaus Von Der Mark

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

During endochondral ossification hypertrophic chondrocytes in the growth plate of fetal long bones, ribs and vertebrae play a key role in preparing growth plate cartilage for replacement by bone. In order to establish a reporter gene mouse to facilitate functional analysis of genes expressed in hypertrophic chondrocytes in this process, Col10a1- BAC reporter gene mouse lines were established expressing LacZ specifically in hypertrophic cartilage under the control of the complete Col10a1 gene. For this purpose, a bacterial artificial chromosome (BAC RP23-192A7) containing the entire murine Col10a1 gene together with 200 kb flanking sequences was modified by inserting a LacZ-Neo cassette into the second exon of Col10a1 by homologous recombination in E. coli. Transgenic mice containing between one and seven transgene copies were generated by injection of the purified BAC-Col10a1- lLacZ DNA. X-gal staining of newborns and embryos revealed strong and robust LacZ activity exclusively in hypertrophic cartilage of the fetal and neonatal skeleton of the transgenic offspring. This indicates that expression of the reporter gene in its proper genomic context in the BAC Col10a1 environment is independent of the integration site and reflects authentic Col10a1 expression in vivo. The Col10a1 specific BAC recombination vector described here will enable the specific analysis of effector gene functions in hypertrophic cartilage during skeletal development, endochondral ossification, and fracture callus healing.

Original languageEnglish
Pages (from-to)183-194
Number of pages12
JournalHistochemistry and Cell Biology
Volume127
Issue number2
DOIs
Publication statusPublished - Feb 2007

Fingerprint

Chondrocytes
genes
Transgenic Mice
Cartilage
mice
Genes
Reporter Genes
cartilage
Growth Plate
Osteogenesis
Bacterial Artificial Chromosomes
Bone and Bones
Fracture Healing
Homologous Recombination
Bony Callus
Ribs
bones
Transgenes
Skeleton
Bone

Keywords

  • BAC
  • Collagen X
  • Homologous recombination
  • Hypertrophic cartilage
  • Transgenic mouse

ASJC Scopus subject areas

  • Cell Biology
  • Instrumentation

Cite this

BAC constructs in transgenic reporter mouse lines control efficient and specific LacZ expression in hypertrophic chondrocytes under the complete Col10a1 promoter. / Gebhard, Sonja; Hattori, Takako; Bauer, Eva; Bösl, Michael R.; Schlund, Britta; Pöschl, Ernst; Adam, Nadia; De Crombrugghe, Benoit; Von Der Mark, Klaus.

In: Histochemistry and Cell Biology, Vol. 127, No. 2, 02.2007, p. 183-194.

Research output: Contribution to journalArticle

Gebhard, Sonja ; Hattori, Takako ; Bauer, Eva ; Bösl, Michael R. ; Schlund, Britta ; Pöschl, Ernst ; Adam, Nadia ; De Crombrugghe, Benoit ; Von Der Mark, Klaus. / BAC constructs in transgenic reporter mouse lines control efficient and specific LacZ expression in hypertrophic chondrocytes under the complete Col10a1 promoter. In: Histochemistry and Cell Biology. 2007 ; Vol. 127, No. 2. pp. 183-194.
@article{095a39061fbf4ff19bc9aa9bef2fc037,
title = "BAC constructs in transgenic reporter mouse lines control efficient and specific LacZ expression in hypertrophic chondrocytes under the complete Col10a1 promoter",
abstract = "During endochondral ossification hypertrophic chondrocytes in the growth plate of fetal long bones, ribs and vertebrae play a key role in preparing growth plate cartilage for replacement by bone. In order to establish a reporter gene mouse to facilitate functional analysis of genes expressed in hypertrophic chondrocytes in this process, Col10a1- BAC reporter gene mouse lines were established expressing LacZ specifically in hypertrophic cartilage under the control of the complete Col10a1 gene. For this purpose, a bacterial artificial chromosome (BAC RP23-192A7) containing the entire murine Col10a1 gene together with 200 kb flanking sequences was modified by inserting a LacZ-Neo cassette into the second exon of Col10a1 by homologous recombination in E. coli. Transgenic mice containing between one and seven transgene copies were generated by injection of the purified BAC-Col10a1- lLacZ DNA. X-gal staining of newborns and embryos revealed strong and robust LacZ activity exclusively in hypertrophic cartilage of the fetal and neonatal skeleton of the transgenic offspring. This indicates that expression of the reporter gene in its proper genomic context in the BAC Col10a1 environment is independent of the integration site and reflects authentic Col10a1 expression in vivo. The Col10a1 specific BAC recombination vector described here will enable the specific analysis of effector gene functions in hypertrophic cartilage during skeletal development, endochondral ossification, and fracture callus healing.",
keywords = "BAC, Collagen X, Homologous recombination, Hypertrophic cartilage, Transgenic mouse",
author = "Sonja Gebhard and Takako Hattori and Eva Bauer and B{\"o}sl, {Michael R.} and Britta Schlund and Ernst P{\"o}schl and Nadia Adam and {De Crombrugghe}, Benoit and {Von Der Mark}, Klaus",
year = "2007",
month = "2",
doi = "10.1007/s00418-006-0236-8",
language = "English",
volume = "127",
pages = "183--194",
journal = "Histochemistry and Cell Biology",
issn = "0948-6143",
publisher = "Springer Verlag",
number = "2",

}

TY - JOUR

T1 - BAC constructs in transgenic reporter mouse lines control efficient and specific LacZ expression in hypertrophic chondrocytes under the complete Col10a1 promoter

AU - Gebhard, Sonja

AU - Hattori, Takako

AU - Bauer, Eva

AU - Bösl, Michael R.

AU - Schlund, Britta

AU - Pöschl, Ernst

AU - Adam, Nadia

AU - De Crombrugghe, Benoit

AU - Von Der Mark, Klaus

PY - 2007/2

Y1 - 2007/2

N2 - During endochondral ossification hypertrophic chondrocytes in the growth plate of fetal long bones, ribs and vertebrae play a key role in preparing growth plate cartilage for replacement by bone. In order to establish a reporter gene mouse to facilitate functional analysis of genes expressed in hypertrophic chondrocytes in this process, Col10a1- BAC reporter gene mouse lines were established expressing LacZ specifically in hypertrophic cartilage under the control of the complete Col10a1 gene. For this purpose, a bacterial artificial chromosome (BAC RP23-192A7) containing the entire murine Col10a1 gene together with 200 kb flanking sequences was modified by inserting a LacZ-Neo cassette into the second exon of Col10a1 by homologous recombination in E. coli. Transgenic mice containing between one and seven transgene copies were generated by injection of the purified BAC-Col10a1- lLacZ DNA. X-gal staining of newborns and embryos revealed strong and robust LacZ activity exclusively in hypertrophic cartilage of the fetal and neonatal skeleton of the transgenic offspring. This indicates that expression of the reporter gene in its proper genomic context in the BAC Col10a1 environment is independent of the integration site and reflects authentic Col10a1 expression in vivo. The Col10a1 specific BAC recombination vector described here will enable the specific analysis of effector gene functions in hypertrophic cartilage during skeletal development, endochondral ossification, and fracture callus healing.

AB - During endochondral ossification hypertrophic chondrocytes in the growth plate of fetal long bones, ribs and vertebrae play a key role in preparing growth plate cartilage for replacement by bone. In order to establish a reporter gene mouse to facilitate functional analysis of genes expressed in hypertrophic chondrocytes in this process, Col10a1- BAC reporter gene mouse lines were established expressing LacZ specifically in hypertrophic cartilage under the control of the complete Col10a1 gene. For this purpose, a bacterial artificial chromosome (BAC RP23-192A7) containing the entire murine Col10a1 gene together with 200 kb flanking sequences was modified by inserting a LacZ-Neo cassette into the second exon of Col10a1 by homologous recombination in E. coli. Transgenic mice containing between one and seven transgene copies were generated by injection of the purified BAC-Col10a1- lLacZ DNA. X-gal staining of newborns and embryos revealed strong and robust LacZ activity exclusively in hypertrophic cartilage of the fetal and neonatal skeleton of the transgenic offspring. This indicates that expression of the reporter gene in its proper genomic context in the BAC Col10a1 environment is independent of the integration site and reflects authentic Col10a1 expression in vivo. The Col10a1 specific BAC recombination vector described here will enable the specific analysis of effector gene functions in hypertrophic cartilage during skeletal development, endochondral ossification, and fracture callus healing.

KW - BAC

KW - Collagen X

KW - Homologous recombination

KW - Hypertrophic cartilage

KW - Transgenic mouse

UR - http://www.scopus.com/inward/record.url?scp=33846413247&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846413247&partnerID=8YFLogxK

U2 - 10.1007/s00418-006-0236-8

DO - 10.1007/s00418-006-0236-8

M3 - Article

C2 - 17051351

AN - SCOPUS:33846413247

VL - 127

SP - 183

EP - 194

JO - Histochemistry and Cell Biology

JF - Histochemistry and Cell Biology

SN - 0948-6143

IS - 2

ER -