Reference Report

Reference
Citation	Bischof, J., Maeda, R.K., Hediger, M., Karch, F., Basler, K. (2007). An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrases. Proc. Natl. Acad. Sci. U.S.A. 104(9): 3312--3317. (Export to RIS)
FlyBase ID	FBrf0192974
Publication Type	Research paper
PubMed ID	17360644
PubMed Abstract	Germ-line transformation via transposable elements is a powerful tool to study gene function in Drosophila melanogaster. However, some inherent characteristics of transposon-mediated transgenesis limit its use for transgene analysis. Here, we circumvent these limitations by optimizing a phiC31-based integration system. We generated a collection of lines with precisely mapped attP sites that allow the insertion of transgenes into many different predetermined intergenic locations throughout the fly genome. By using regulatory elements of the nanos and vasa genes, we established endogenous sources of the phiC31 integrase, eliminating the difficulties of coinjecting integrase mRNA and raising the transformation efficiency. Moreover, to discriminate between specific and rare nonspecific integration events, a white gene-based reconstitution system was generated that enables visual selection for precise attP targeting. Finally, we demonstrate that our chromosomal attP sites can be modified in situ, extending their scope while retaining their properties as landing sites. The efficiency, ease-of-use, and versatility obtained here with the phiC31-based integration system represents an important advance in transgenesis and opens up the possibility of systematic, high-throughput screening of large cDNA sets and regulatory elements.
DOI	10.1073/pnas.0611511104
Related Publication(s)
Supplementary material	SI Table 4. [FBrf0195884] SI Text. [FBrf0198668] SI Figure 4. [FBrf0198669] SI Figure 5. [FBrf0198670]
Recent Updates
Description	What does this section display? This section contains items that were added to this record for each release. It currently only tracks new links between this FlyBase report and other FlyBase data classes (e.g. genes, references, stocks) or controlled vocabulary terms (e.g. GO, anatomy terms). What does this section not display? This section does not currently display links that were removed or gene model changes.
Update Feed	Click the icon below to subscribe to this FlyBase record and receive updates automatically through your feed reader.
FB2013_03
FB2013_02
All updates	Click here to see a list of all updates to this record from FB2010_08 and on.
Associated Information
Comments
Associated Files
Other Information
Secondary IDs
Language of Publication	English
Additional Languages of Abstract
Also Published As
Parent Publication
Publication Type	Journal
Abbreviation	Proc. Natl. Acad. Sci. U.S.A.
Title	Proceedings of the National Academy of Sciences of the United States of America
Publication Year	1915-
ISBN/ISSN	0027-8424
Data from Reference
Alleles (12)
	Avic\GFP^E.3xP3 Disc\RFP^3xP3.PB Ecol\lacZ^T:SV40\nls2 phiC31\int^nos.PB phiC31\int^{nos.T:SV40\nls2} phiC31\int^vas.PB phiC31\int^{vas.T:SV40\nls2} Scer\GAL4^ap-md544 Scer\GAL4^bi-md653 w^+mC w^Ex3-6 w^P-Ex2
Constructs (18)
	M{3xP3-RFP.attP'} M{3xP3-RFP.attP} M{loxP.attP} M{nos-int.B} M{nos-int.NLS} M{UAST-lacZ.B} M{vas-int.B} M{vas-int.NLS} M{w^Ex3-6.attP} M{w^Ex3-6.w^P-Ex2} p3xP3-EGFP.nos-int.attB p3xP3-EGFP.nos-int.NLS.attB p3xP3-EGFP.vas-int.attB p3xP3-EGFP.vas-int.NLS.attB p3xP3-RFP.wEx3-6.attP.attB P{nos-phiC31\int.NLS} pUAS-lacZattB pw^P-Ex2UASTattB
Genes (5)
	Avic\GFP Disc\RFP phiC31\int Scer\GAL4 w
Insertions (18)
	M{3xP3-RFP.attP}ZH-2A M{3xP3-RFP.attP}ZH-86Fb M{3xP3-RFP.attP}ZH-102D M{loxP.attP}ZH-86Fb M{nos-int.B}ZH-86Fb M{nos-int.B}ZH-102D M{nos-int.NLS}ZH-86Fb M{nos-int.NLS}ZH-102D M{UAST-lacZ.B}ZH-86Fb M{vas-int.B}ZH-86Fb M{vas-int.B}ZH-102D M{vas-int.NLS}ZH-86Fb M{vas-int.NLS}ZH-102D M{w^Ex3-6.attP}ZH-86Fb M{w^Ex3-6.w^P-Ex2}ZH-86Fb P{GawB}ap^md544 P{GawB}bi^md653 P{nos-phiC31\int.NLS}X
Natural transposons (2)
	Dmau\mariner P-element