Context dependent function of APPb enhancer identified using enhancer trap-containing BACs as transgenes in zebrafish

Leighcraft A. Shakes, Tennison L. Malcolm, Kevin L. Allen, Supriyo De, Ken R. Harewood, Pradeep K. Chatterjee

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

An enhancer within intron 1 of the amyloid precursor protein gene (APPb) of zebrafish is identified functionally using a novel approach. Bacterial artificial chromosomes (BACs) were retrofitted with enhancer traps, and expressed as transgenes in zebrafish. Expression from both transient assays and stable lines were used for analysis. Although the enhancer was active in specific nonneural cells of the notochord when placed with APPb gene promoter proximal elements its function was restricted to, and absolutely required for, specific expression in neurons when juxtaposed with additional far-upstream promoter elements of the gene. We demonstrate that expression of green fluorescent protein fluorescence resembling the tissue distribution of APPb mRNA requires both the intron 1 enhancer and ∼28 kb of DNA upstream of the gene. The results indicate that tissue-specificity of an isolated enhancer may be quite different from that in the context of its own gene. Using this enhancer and upstream sequence, polymorphic variants of APPb can now more closely recapitulate the endogenous pattern and regulation of APPb expression in animal models for Alzheimer's disease. The methodology should help functionally map multiple noncontiguous regulatory elements in BACs with or without gene-coding sequences.

Original languageEnglish (US)
Pages (from-to)6237-6248
Number of pages12
JournalNucleic Acids Research
Volume36
Issue number19
DOIs
StatePublished - Nov 19 2008

Fingerprint

Bacterial Artificial Chromosomes
Zebrafish
Transgenes
Genes
Introns
Notochord
Organ Specificity
Amyloid beta-Protein Precursor
Tissue Distribution
Green Fluorescent Proteins
Alzheimer Disease
Animal Models
Fluorescence
Neurons
Messenger RNA
DNA

ASJC Scopus subject areas

  • Genetics

Cite this

Context dependent function of APPb enhancer identified using enhancer trap-containing BACs as transgenes in zebrafish. / Shakes, Leighcraft A.; Malcolm, Tennison L.; Allen, Kevin L.; De, Supriyo; Harewood, Ken R.; Chatterjee, Pradeep K.

In: Nucleic Acids Research, Vol. 36, No. 19, 19.11.2008, p. 6237-6248.

Research output: Contribution to journalArticle

Shakes, Leighcraft A. ; Malcolm, Tennison L. ; Allen, Kevin L. ; De, Supriyo ; Harewood, Ken R. ; Chatterjee, Pradeep K. / Context dependent function of APPb enhancer identified using enhancer trap-containing BACs as transgenes in zebrafish. In: Nucleic Acids Research. 2008 ; Vol. 36, No. 19. pp. 6237-6248.
@article{aaf6e35f39a34f76993a22fc4c4208bf,
title = "Context dependent function of APPb enhancer identified using enhancer trap-containing BACs as transgenes in zebrafish",
abstract = "An enhancer within intron 1 of the amyloid precursor protein gene (APPb) of zebrafish is identified functionally using a novel approach. Bacterial artificial chromosomes (BACs) were retrofitted with enhancer traps, and expressed as transgenes in zebrafish. Expression from both transient assays and stable lines were used for analysis. Although the enhancer was active in specific nonneural cells of the notochord when placed with APPb gene promoter proximal elements its function was restricted to, and absolutely required for, specific expression in neurons when juxtaposed with additional far-upstream promoter elements of the gene. We demonstrate that expression of green fluorescent protein fluorescence resembling the tissue distribution of APPb mRNA requires both the intron 1 enhancer and ∼28 kb of DNA upstream of the gene. The results indicate that tissue-specificity of an isolated enhancer may be quite different from that in the context of its own gene. Using this enhancer and upstream sequence, polymorphic variants of APPb can now more closely recapitulate the endogenous pattern and regulation of APPb expression in animal models for Alzheimer's disease. The methodology should help functionally map multiple noncontiguous regulatory elements in BACs with or without gene-coding sequences.",
author = "Shakes, {Leighcraft A.} and Malcolm, {Tennison L.} and Allen, {Kevin L.} and Supriyo De and Harewood, {Ken R.} and Chatterjee, {Pradeep K.}",
year = "2008",
month = "11",
day = "19",
doi = "10.1093/nar/gkn628",
language = "English (US)",
volume = "36",
pages = "6237--6248",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "19",

}

TY - JOUR

T1 - Context dependent function of APPb enhancer identified using enhancer trap-containing BACs as transgenes in zebrafish

AU - Shakes, Leighcraft A.

AU - Malcolm, Tennison L.

AU - Allen, Kevin L.

AU - De, Supriyo

AU - Harewood, Ken R.

AU - Chatterjee, Pradeep K.

PY - 2008/11/19

Y1 - 2008/11/19

N2 - An enhancer within intron 1 of the amyloid precursor protein gene (APPb) of zebrafish is identified functionally using a novel approach. Bacterial artificial chromosomes (BACs) were retrofitted with enhancer traps, and expressed as transgenes in zebrafish. Expression from both transient assays and stable lines were used for analysis. Although the enhancer was active in specific nonneural cells of the notochord when placed with APPb gene promoter proximal elements its function was restricted to, and absolutely required for, specific expression in neurons when juxtaposed with additional far-upstream promoter elements of the gene. We demonstrate that expression of green fluorescent protein fluorescence resembling the tissue distribution of APPb mRNA requires both the intron 1 enhancer and ∼28 kb of DNA upstream of the gene. The results indicate that tissue-specificity of an isolated enhancer may be quite different from that in the context of its own gene. Using this enhancer and upstream sequence, polymorphic variants of APPb can now more closely recapitulate the endogenous pattern and regulation of APPb expression in animal models for Alzheimer's disease. The methodology should help functionally map multiple noncontiguous regulatory elements in BACs with or without gene-coding sequences.

AB - An enhancer within intron 1 of the amyloid precursor protein gene (APPb) of zebrafish is identified functionally using a novel approach. Bacterial artificial chromosomes (BACs) were retrofitted with enhancer traps, and expressed as transgenes in zebrafish. Expression from both transient assays and stable lines were used for analysis. Although the enhancer was active in specific nonneural cells of the notochord when placed with APPb gene promoter proximal elements its function was restricted to, and absolutely required for, specific expression in neurons when juxtaposed with additional far-upstream promoter elements of the gene. We demonstrate that expression of green fluorescent protein fluorescence resembling the tissue distribution of APPb mRNA requires both the intron 1 enhancer and ∼28 kb of DNA upstream of the gene. The results indicate that tissue-specificity of an isolated enhancer may be quite different from that in the context of its own gene. Using this enhancer and upstream sequence, polymorphic variants of APPb can now more closely recapitulate the endogenous pattern and regulation of APPb expression in animal models for Alzheimer's disease. The methodology should help functionally map multiple noncontiguous regulatory elements in BACs with or without gene-coding sequences.

UR - http://www.scopus.com/inward/record.url?scp=56049108103&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=56049108103&partnerID=8YFLogxK

U2 - 10.1093/nar/gkn628

DO - 10.1093/nar/gkn628

M3 - Article

C2 - 18832376

AN - SCOPUS:56049108103

VL - 36

SP - 6237

EP - 6248

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 19

ER -