PUBLICATION

Isolation of Novel CreERT2-Driver Lines in Zebrafish Using an Unbiased Gene Trap Approach

Authors

Jungke, P., Hammer, J., Hans, S., Brand, M.

ID

ZDB-PUB-150618-1

Date

2015

Source

PLoS One 10: e0129072 (Journal)

Registered Authors

Brand, Michael, Hans, Stefan, Jungke, Peggy

Keywords

none

MeSH Terms

Gene Expression Regulation, Developmental
Luminescent Proteins/genetics
Luminescent Proteins/metabolism
Genetic Vectors/chemistry*
Genetic Vectors/metabolism
Animals, Genetically Modified
Green Fluorescent Proteins/genetics
Green Fluorescent Proteins/metabolism
Embryo, Nonmammalian
Genes, Reporter
Zebrafish/embryology
Zebrafish/genetics*
Zebrafish/metabolism
Animals
Integrases/genetics*
Integrases/metabolism
Microinjections
Transgenes
Zygote/growth & development
Zygote/metabolism*
Promoter Regions, Genetic

(all 21)

PubMed

26083735 Full text @ PLoS One

Abstract

Gene manipulation using the Cre/loxP-recombinase system has been successfully employed in zebrafish to study gene functions and lineage relationships. Recently, gene trapping approaches have been applied to produce large collections of transgenic fish expressing conditional alleles in various tissues. However, the limited number of available cell- and tissue-specific Cre/CreERT2-driver lines still constrains widespread application in this model organism. To enlarge the pool of existing CreERT2-driver lines, we performed a genome-wide gene trap screen using a Tol2-based mCherry-T2a-CreERT2 (mCT2aC) gene trap vector. This cassette consists of a splice acceptor and a mCherry-tagged variant of CreERT2 which enables simultaneous labeling of the trapping event, as well as CreERT2 expression from the endogenous promoter. Using this strategy, we generated 27 novel functional CreERT2-driver lines expressing in a cell- and tissue-specific manner during development and adulthood. This study summarizes the analysis of the generated CreERT2-driver lines with respect to functionality, expression, integration, as well as associated phenotypes. Our results significantly enlarge the existing pool of CreERT2-driver lines in zebrafish and combined with Cre-dependent effector lines, the new CreERT2-driver lines will be important tools to manipulate the zebrafish genome.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
EGFP	baz2ba^tud21Gt; tud107Tg	heat shock, chemical treatment: tamoxifen	Prim-5		neural tube	IFL	Fig. 3
EGFP	ccdc102a^tud29Gt; tud107Tg	heat shock, chemical treatment: tamoxifen	Prim-5		anatomical structure	IFL	Fig. 3
EGFP	ccdc102a^tud29Gt; tud107Tg	heat shock	Day 4	Not Detected	anatomical structure	IFL	Fig. 4
EGFP	ccdc102a^tud29Gt; tud107Tg	heat shock, chemical treatment: afimoxifene	Day 4		head	IFL	Fig. 4
EGFP	cdc42ep4b^tud16Gt; tud107Tg	heat shock, chemical treatment: tamoxifen	Prim-5		anatomical structure	IFL	Fig. 3
EGFP	epha7^tud27Gt; tud107Tg	heat shock, chemical treatment: tamoxifen	Prim-5		anatomical structure	IFL	Fig. 3
EGFP	hoxb1b^tud17Gt; tud107Tg	heat shock, chemical treatment: tamoxifen	Prim-5		spinal cord	IFL	Fig. 3
EGFP	hoxb1b^tud17Gt; tud107Tg	heat shock, chemical treatment: tamoxifen	Prim-5		telencephalon	IFL	Fig. 3
EGFP	klhl17^tud15Gt; tud107Tg	heat shock, chemical treatment: tamoxifen	Prim-5		anatomical structure	IFL	Fig. 3
EGFP	mapre1a^tud12Gt; tud107Tg	heat shock, chemical treatment: tamoxifen	Prim-5		anatomical structure	IFL	Fig. 3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
otx1^{tud37Gt/tud37Gt} (AB)	standard conditions	Protruding-mouth	eye morphology, abnormal	Fig. 9
otx1^{tud37Gt/tud37Gt} (AB)	standard conditions	Protruding-mouth	forebrain morphology, abnormal	Fig. 9
otx1^{tud37Gt/tud37Gt} (AB)	standard conditions	Protruding-mouth	heart morphology, abnormal	Fig. 9
otx1^{tud37Gt/tud37Gt} (AB)	standard conditions	Protruding-mouth	inner ear morphology, abnormal	Fig. 9
otx1^{tud37Gt/tud37Gt} (AB)	standard conditions	Protruding-mouth	midbrain morphology, abnormal	Fig. 9
otx1^{tud37Gt/tud37Gt} (AB)	standard conditions	Protruding-mouth	whole organism anterior-posterior axis bent, abnormal	Fig. 9

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
tud11Gt	Gt(SA1-mCT2aC)	Transgenic Insertion	ptk2aa
tud12Gt	Gt(SA1-mCT2aC)	Transgenic Insertion	mapre1a
tud13Gt	Gt(SA1-mCT2aC)	Transgenic Insertion	pvalb1
tud14Gt	Gt(SA1-mCT2aC)	Transgenic Insertion
tud15Gt	Gt(SA1-mCT2aC)	Transgenic Insertion	klhl17
tud16Gt	Gt(SA1-mCT2aC)	Transgenic Insertion	cdc42ep4b
tud17Gt	Gt(SA1-mCT2aC)	Transgenic Insertion	hoxb1b
tud18Gt	Gt(SA1-mCT2aC)	Transgenic Insertion	msx3
tud19Gt	Gt(SA1-mCT2aC)	Transgenic Insertion
tud20Gt	Gt(SA1-mCT2aC)	Transgenic Insertion	si:ch73-248e17.1

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Human Disease / Model

Sequence Targeting Reagents

Fish

Fish
baz2ba^tud21Gt; tud107Tg
ccdc102a^tud29Gt; tud107Tg
cdc42ep4b^tud16Gt; tud107Tg
epha7^tud27Gt (AB)
epha7^tud27Gt; tud107Tg
hoxb1b^tud17Gt (AB)
hoxb1b^tud17Gt; tud107Tg
klhl17^tud15Gt; tud107Tg
mapre1a^tud12Gt; tud107Tg
msx3^tud18Gt; tud107Tg

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
Cre	EFG	Cre
DsRed2	EFG	DsRed2
EGFP	EFG	EGFP
mCherry	EFG	mCherry

Mapping

Jungke et al., 2015 ZDB-PUB-150618-1 PMID:26083735

Isolation of Novel CreERT2-Driver Lines in Zebrafish Using an Unbiased Gene Trap Approach

Jungke et al., 2015

ZDB-PUB-150618-1
PMID:26083735