An allelic series for the leptin receptor gene generated by CRE and FLP recombinase

Julie E. McMinn, Shun Mei Liu, Ioannis Dragatsis, Paula Dietrich, Thomas Ludwig, Sandra Eiden, Streamson C. Chua

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Body weight regulation is mediated through several major signaling pathways, some of which have been delineated by positional cloning of spontaneous genetic mutations in mice. Leprdb/db mice are obese due to a defect in the signaling portion of the leptin receptor, which has led to extensive study of this highly conserved system over the past several years. We have created an allelic series at Lepr for the further examination of LEPR signaling phenotypes using both the FLP/frt and CRE/foxP systems. By inserting a frt-PGK-neo-frt sequence in Lepr intron 16, we have generated a conditional gene repair Lepr allele (Lepr-neo) that elicits morbid obesity, diabetes, and infertility in homozygous mice, recapitulating the obesity syndrome of Lep db/db mice. Thus, in vivo excision of the PGK-neo cassette with a FLP recombinase transgene restores the lean and fertile phenotype to Lepr flox/flox mice. In the same construct, we have also inserted loxP sites that flank Lepr coding exon 17, a region that encodes a JAK docking site required for STAT3 signaling. CRE-mediated excision of Lepr coding exon 17 from Lepr with a frameshift in subsequent exons results in a syndrome of obesity, diabetes, and infertility in LeprΔ17/Δ17 mice, which is indistinguishable from Leprneo/neo and Leprdb/db mice. We conclude that suppression of Lepr gene expression by PGK-neo is phenotypically equivalent to deletion of the Lepr signaling motifs, and therefore the Lepr neo/neo mouse may be used to investigate conditional gene repair of Lepr signaling deficiency.

Original languageEnglish (US)
Pages (from-to)677-685
Number of pages9
JournalMammalian Genome
Volume15
Issue number9
DOIs
StatePublished - Sep 1 2004

Fingerprint

Leptin Receptors
Genes
Exons
Infertility
Obesity
Phenotype
Obese Mice
Morbid Obesity
FLP recombinase
Transgenes
Introns
Organism Cloning
Alleles
Body Weight
Gene Expression
Mutation

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

An allelic series for the leptin receptor gene generated by CRE and FLP recombinase. / McMinn, Julie E.; Liu, Shun Mei; Dragatsis, Ioannis; Dietrich, Paula; Ludwig, Thomas; Eiden, Sandra; Chua, Streamson C.

In: Mammalian Genome, Vol. 15, No. 9, 01.09.2004, p. 677-685.

Research output: Contribution to journalArticle

McMinn, Julie E. ; Liu, Shun Mei ; Dragatsis, Ioannis ; Dietrich, Paula ; Ludwig, Thomas ; Eiden, Sandra ; Chua, Streamson C. / An allelic series for the leptin receptor gene generated by CRE and FLP recombinase. In: Mammalian Genome. 2004 ; Vol. 15, No. 9. pp. 677-685.
@article{751fe882c9fa48ceb9d66650bb6fa6ff,
title = "An allelic series for the leptin receptor gene generated by CRE and FLP recombinase",
abstract = "Body weight regulation is mediated through several major signaling pathways, some of which have been delineated by positional cloning of spontaneous genetic mutations in mice. Leprdb/db mice are obese due to a defect in the signaling portion of the leptin receptor, which has led to extensive study of this highly conserved system over the past several years. We have created an allelic series at Lepr for the further examination of LEPR signaling phenotypes using both the FLP/frt and CRE/foxP systems. By inserting a frt-PGK-neo-frt sequence in Lepr intron 16, we have generated a conditional gene repair Lepr allele (Lepr-neo) that elicits morbid obesity, diabetes, and infertility in homozygous mice, recapitulating the obesity syndrome of Lep db/db mice. Thus, in vivo excision of the PGK-neo cassette with a FLP recombinase transgene restores the lean and fertile phenotype to Lepr flox/flox mice. In the same construct, we have also inserted loxP sites that flank Lepr coding exon 17, a region that encodes a JAK docking site required for STAT3 signaling. CRE-mediated excision of Lepr coding exon 17 from Lepr with a frameshift in subsequent exons results in a syndrome of obesity, diabetes, and infertility in LeprΔ17/Δ17 mice, which is indistinguishable from Leprneo/neo and Leprdb/db mice. We conclude that suppression of Lepr gene expression by PGK-neo is phenotypically equivalent to deletion of the Lepr signaling motifs, and therefore the Lepr neo/neo mouse may be used to investigate conditional gene repair of Lepr signaling deficiency.",
author = "McMinn, {Julie E.} and Liu, {Shun Mei} and Ioannis Dragatsis and Paula Dietrich and Thomas Ludwig and Sandra Eiden and Chua, {Streamson C.}",
year = "2004",
month = "9",
day = "1",
doi = "10.1007/s00335-004-2340-1",
language = "English (US)",
volume = "15",
pages = "677--685",
journal = "Mammalian Genome",
issn = "0938-8990",
publisher = "Springer New York",
number = "9",

}

TY - JOUR

T1 - An allelic series for the leptin receptor gene generated by CRE and FLP recombinase

AU - McMinn, Julie E.

AU - Liu, Shun Mei

AU - Dragatsis, Ioannis

AU - Dietrich, Paula

AU - Ludwig, Thomas

AU - Eiden, Sandra

AU - Chua, Streamson C.

PY - 2004/9/1

Y1 - 2004/9/1

N2 - Body weight regulation is mediated through several major signaling pathways, some of which have been delineated by positional cloning of spontaneous genetic mutations in mice. Leprdb/db mice are obese due to a defect in the signaling portion of the leptin receptor, which has led to extensive study of this highly conserved system over the past several years. We have created an allelic series at Lepr for the further examination of LEPR signaling phenotypes using both the FLP/frt and CRE/foxP systems. By inserting a frt-PGK-neo-frt sequence in Lepr intron 16, we have generated a conditional gene repair Lepr allele (Lepr-neo) that elicits morbid obesity, diabetes, and infertility in homozygous mice, recapitulating the obesity syndrome of Lep db/db mice. Thus, in vivo excision of the PGK-neo cassette with a FLP recombinase transgene restores the lean and fertile phenotype to Lepr flox/flox mice. In the same construct, we have also inserted loxP sites that flank Lepr coding exon 17, a region that encodes a JAK docking site required for STAT3 signaling. CRE-mediated excision of Lepr coding exon 17 from Lepr with a frameshift in subsequent exons results in a syndrome of obesity, diabetes, and infertility in LeprΔ17/Δ17 mice, which is indistinguishable from Leprneo/neo and Leprdb/db mice. We conclude that suppression of Lepr gene expression by PGK-neo is phenotypically equivalent to deletion of the Lepr signaling motifs, and therefore the Lepr neo/neo mouse may be used to investigate conditional gene repair of Lepr signaling deficiency.

AB - Body weight regulation is mediated through several major signaling pathways, some of which have been delineated by positional cloning of spontaneous genetic mutations in mice. Leprdb/db mice are obese due to a defect in the signaling portion of the leptin receptor, which has led to extensive study of this highly conserved system over the past several years. We have created an allelic series at Lepr for the further examination of LEPR signaling phenotypes using both the FLP/frt and CRE/foxP systems. By inserting a frt-PGK-neo-frt sequence in Lepr intron 16, we have generated a conditional gene repair Lepr allele (Lepr-neo) that elicits morbid obesity, diabetes, and infertility in homozygous mice, recapitulating the obesity syndrome of Lep db/db mice. Thus, in vivo excision of the PGK-neo cassette with a FLP recombinase transgene restores the lean and fertile phenotype to Lepr flox/flox mice. In the same construct, we have also inserted loxP sites that flank Lepr coding exon 17, a region that encodes a JAK docking site required for STAT3 signaling. CRE-mediated excision of Lepr coding exon 17 from Lepr with a frameshift in subsequent exons results in a syndrome of obesity, diabetes, and infertility in LeprΔ17/Δ17 mice, which is indistinguishable from Leprneo/neo and Leprdb/db mice. We conclude that suppression of Lepr gene expression by PGK-neo is phenotypically equivalent to deletion of the Lepr signaling motifs, and therefore the Lepr neo/neo mouse may be used to investigate conditional gene repair of Lepr signaling deficiency.

UR - http://www.scopus.com/inward/record.url?scp=4344713706&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4344713706&partnerID=8YFLogxK

U2 - 10.1007/s00335-004-2340-1

DO - 10.1007/s00335-004-2340-1

M3 - Article

VL - 15

SP - 677

EP - 685

JO - Mammalian Genome

JF - Mammalian Genome

SN - 0938-8990

IS - 9

ER -