A new and simple approach for genotyping Alzheimer's disease presenilin-1 mutant knock-in mice

Vanessa Gautheron, Alexandra Auffret, Mark P. Mattson, Jean Mariani, Béatrice Vernet der Garabedian

Research output: Contribution to journalArticle

Abstract

The use of transgenic mice expressing point mutations demands that the detection of the different alleles is efficient and reliable. In addition, the multiplication of transgenes included in mouse models of human disease underlines the importance of correct controls and the fact that investigators need an accurate and rapid genotyping of the littermates generated. In this study, we demonstrate a powerful alternative for genotyping using presenilin-1 mutant knock-in (PS1M146KI) mice as an example. Mutations in the presenilin-1 (PS1) gene are causally linked to many cases of early-onset inherited Alzheimer's disease (AD). PS1M146VKI mice that express the PS1M146V targeted allele at normal physiological levels and triple-transgenic model (3×Tg-AD) derived from homozygous PS1M146VKI mice were generated to study the pathogenesis of AD. Genotyping PS1M146VKI line requires many steps and thus a large quantity of DNA. In PS1M146VKI mice, only three nucleotides are modified in the gene. Here we show that this small mutated DNA sequence can affect its secondary structure resulting in altered mobility that can be easily detected on a polyacrylamide gel, by the single-strand conformation polymorphism (SSCP) technique. Our results demonstrate that SSCP is a simple, accurate, repeatable and efficient method for the routine genotyping of this current AD model. This method could be easily applied to other transgenic mice.

Original languageEnglish (US)
Pages (from-to)235-240
Number of pages6
JournalJournal of Neuroscience Methods
Volume181
Issue number2
DOIs
StatePublished - Jul 30 2009
Externally publishedYes

Fingerprint

Presenilin-1
Alzheimer Disease
Transgenic Mice
Alleles
Transgenes
Point Mutation
Genes
Nucleotides
Research Personnel
Alzheimer disease type 1
Mutation
DNA

Keywords

  • Alzheimer's disease
  • Genotyping
  • Knock-in
  • Presenilin-1
  • SSCP

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

A new and simple approach for genotyping Alzheimer's disease presenilin-1 mutant knock-in mice. / Gautheron, Vanessa; Auffret, Alexandra; Mattson, Mark P.; Mariani, Jean; Garabedian, Béatrice Vernet der.

In: Journal of Neuroscience Methods, Vol. 181, No. 2, 30.07.2009, p. 235-240.

Research output: Contribution to journalArticle

Gautheron, Vanessa ; Auffret, Alexandra ; Mattson, Mark P. ; Mariani, Jean ; Garabedian, Béatrice Vernet der. / A new and simple approach for genotyping Alzheimer's disease presenilin-1 mutant knock-in mice. In: Journal of Neuroscience Methods. 2009 ; Vol. 181, No. 2. pp. 235-240.
@article{0e1bfa56887b44808eceb886d56d4d7a,
title = "A new and simple approach for genotyping Alzheimer's disease presenilin-1 mutant knock-in mice",
abstract = "The use of transgenic mice expressing point mutations demands that the detection of the different alleles is efficient and reliable. In addition, the multiplication of transgenes included in mouse models of human disease underlines the importance of correct controls and the fact that investigators need an accurate and rapid genotyping of the littermates generated. In this study, we demonstrate a powerful alternative for genotyping using presenilin-1 mutant knock-in (PS1M146KI) mice as an example. Mutations in the presenilin-1 (PS1) gene are causally linked to many cases of early-onset inherited Alzheimer's disease (AD). PS1M146VKI mice that express the PS1M146V targeted allele at normal physiological levels and triple-transgenic model (3×Tg-AD) derived from homozygous PS1M146VKI mice were generated to study the pathogenesis of AD. Genotyping PS1M146VKI line requires many steps and thus a large quantity of DNA. In PS1M146VKI mice, only three nucleotides are modified in the gene. Here we show that this small mutated DNA sequence can affect its secondary structure resulting in altered mobility that can be easily detected on a polyacrylamide gel, by the single-strand conformation polymorphism (SSCP) technique. Our results demonstrate that SSCP is a simple, accurate, repeatable and efficient method for the routine genotyping of this current AD model. This method could be easily applied to other transgenic mice.",
keywords = "Alzheimer's disease, Genotyping, Knock-in, Presenilin-1, SSCP",
author = "Vanessa Gautheron and Alexandra Auffret and Mattson, {Mark P.} and Jean Mariani and Garabedian, {B{\'e}atrice Vernet der}",
year = "2009",
month = "7",
day = "30",
doi = "10.1016/j.jneumeth.2009.05.009",
language = "English (US)",
volume = "181",
pages = "235--240",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - A new and simple approach for genotyping Alzheimer's disease presenilin-1 mutant knock-in mice

AU - Gautheron, Vanessa

AU - Auffret, Alexandra

AU - Mattson, Mark P.

AU - Mariani, Jean

AU - Garabedian, Béatrice Vernet der

PY - 2009/7/30

Y1 - 2009/7/30

N2 - The use of transgenic mice expressing point mutations demands that the detection of the different alleles is efficient and reliable. In addition, the multiplication of transgenes included in mouse models of human disease underlines the importance of correct controls and the fact that investigators need an accurate and rapid genotyping of the littermates generated. In this study, we demonstrate a powerful alternative for genotyping using presenilin-1 mutant knock-in (PS1M146KI) mice as an example. Mutations in the presenilin-1 (PS1) gene are causally linked to many cases of early-onset inherited Alzheimer's disease (AD). PS1M146VKI mice that express the PS1M146V targeted allele at normal physiological levels and triple-transgenic model (3×Tg-AD) derived from homozygous PS1M146VKI mice were generated to study the pathogenesis of AD. Genotyping PS1M146VKI line requires many steps and thus a large quantity of DNA. In PS1M146VKI mice, only three nucleotides are modified in the gene. Here we show that this small mutated DNA sequence can affect its secondary structure resulting in altered mobility that can be easily detected on a polyacrylamide gel, by the single-strand conformation polymorphism (SSCP) technique. Our results demonstrate that SSCP is a simple, accurate, repeatable and efficient method for the routine genotyping of this current AD model. This method could be easily applied to other transgenic mice.

AB - The use of transgenic mice expressing point mutations demands that the detection of the different alleles is efficient and reliable. In addition, the multiplication of transgenes included in mouse models of human disease underlines the importance of correct controls and the fact that investigators need an accurate and rapid genotyping of the littermates generated. In this study, we demonstrate a powerful alternative for genotyping using presenilin-1 mutant knock-in (PS1M146KI) mice as an example. Mutations in the presenilin-1 (PS1) gene are causally linked to many cases of early-onset inherited Alzheimer's disease (AD). PS1M146VKI mice that express the PS1M146V targeted allele at normal physiological levels and triple-transgenic model (3×Tg-AD) derived from homozygous PS1M146VKI mice were generated to study the pathogenesis of AD. Genotyping PS1M146VKI line requires many steps and thus a large quantity of DNA. In PS1M146VKI mice, only three nucleotides are modified in the gene. Here we show that this small mutated DNA sequence can affect its secondary structure resulting in altered mobility that can be easily detected on a polyacrylamide gel, by the single-strand conformation polymorphism (SSCP) technique. Our results demonstrate that SSCP is a simple, accurate, repeatable and efficient method for the routine genotyping of this current AD model. This method could be easily applied to other transgenic mice.

KW - Alzheimer's disease

KW - Genotyping

KW - Knock-in

KW - Presenilin-1

KW - SSCP

UR - http://www.scopus.com/inward/record.url?scp=67649259010&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67649259010&partnerID=8YFLogxK

U2 - 10.1016/j.jneumeth.2009.05.009

DO - 10.1016/j.jneumeth.2009.05.009

M3 - Article

C2 - 19465058

AN - SCOPUS:67649259010

VL - 181

SP - 235

EP - 240

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

IS - 2

ER -