PhiC31 integrase-mediated transgenesis systems

We use M{vas-int.Dm}ZH-2A as the genomic PhiC31 source if the attP site stock does not contain it. If the screening marker is fluorescence-related, we may use the genomic P{nos-phiC31\int.NLS}X source (non-fluorescence), please let us know if this is the case.

For vermillion⁺ markers, e.g. for creating stable attP-gRNA lines, you may actually choose any of the vermillion⁺ based, non-X, attP lines from these below lists. We may setup the crosses to the y^-v^- line and screen for v⁺. You may get the same v⁺ G₂ transformants as using the v^- based attP lines. This is just regular Plan H or Plan I with extra v⁺ screening. You may also ask us to keep the resulting gRNA lines and have us cross to any of the Cas9 stocks and inject their F1s (additional Plan R) later.

P{CaryP}
Two attP landing sites were created by the Calos group. These two sites were initially created by the traditional P-element system. See below paper for more info.

PhiC31-mediated RMCE
In June, 2006, the Wu lab published a method that targets constructs to predetermined genomic sites using the phiC31 integrase system in conjunction with Recombinase Mediated Cassette Exchange (RMCE).

Please visit the Wu lab for more info:
http://transvection.org

P[acman]
P[acman] system makes use of the combined tools of a conditional amplifiable technique for typical P1 and BAC construct, recombineering, and PhiC31-mediated trangenesis. It has the advantage of delivering large DNA fragments into specific genomic locations with attP site, which were generated by the Ballen lab and markered by yellow. Here we are incorporating the P[acman] system with the work from the Basler group which used endogenous PhiC31 source instead of co-injecting the PhiC31 mRNA.

Please visit the Bellen lab for more info:
http://flypush.imgen.bcm.tmc.edu/lab/index.html

MiMIC
MiMIC is a highly versatile transposon insertion resource for engineering Drosophila melanogaster genes. MiMIC system was developed by the collaboration among the laboratories of Hugo Bellen (Baylor College of Medicine), Roger Hoskins (Lawrence Berkeley National Laboratory) and Allan Spradling (Carnegie Institution of Washington).

Please visit the Bellen lab MiMIC page for more info:
http://flypush.imgen.bcm.tmc.edu/pscreen/technique.html

FlyC31
In FlyC31 system, a library of landing platforms containing attP site throughout the Drosophila genome were created by the Basler group, which are ready for the transgenesis of the construct with attB site. These landing platforms were deliberately designed to be manipulated in vivo by Cre/loxP system after transgenesis to get rid of most of the non-necessary sequences and only keep your gene-of-interest flanking with one loxP and one attL site (We do not provide this Cre/loxP service).

Please visit the FlyC31 website for more info:
http://www.flyc31.org/why_phic31.php

Perrimon Strains
Additional attP landing sites were created by the Perrimon group, and were tested for position effects by luciferase expression on various tissues. See below paper for more info.

Please visit the Perrimon lab website for more info:
http://genetics.med.harvard.edu/~perrimon/

The PhiC31 system and the Gateway System
One of the "Gateway-PhiC31 system", for example, can be found here. Please note the attP1, attP2, attB1, attB2, etc, at the original Murphys Gateway are of different concepts.

Reference:

Groth A, Fish M, Nusse R, Calos MP (2004) Construction of transgenic Drosophila by using the site-specific integrase from phage PhiC31. Genetics 166: 1775

Bateman JR, Lee AM, Wu C-t (2006) Site-specific transformation of Drosophila via PhiC31 integrase-mediated cassette exchange. Genetics 173:769

Venken KJT, He Y, Hoskins RA, Bellen HJ (2006) PhiC31: a BAC transgenic platform for targeted insertion of large DNA fragments in Drosophila melanogaster. Science 314:1747

Bischof J, Maeda RK, Hediger M, Karch F, Basler K (2007) An optimized transgenesis system for Drosophila using germ-line-specific PhiC31 integrases. PNAS 104:3312

Markstein M, Pitsouli C, Villalta C, Celniker SE, Perrimon N (2008) Exploiting position effects and the gypsy retrovirus insulator to engineer precisely expressed transgenes. Nature Genetics 40:476

Currently available Calos strains (P{CaryP} strains) at BestGene Inc.

Currently available Wu strains and Bateman strains (PhiC31-RMCE strains) at BestGene Inc.

Currently available Bellen strains (P[acman] strains) at BestGene Inc.

Currently available Basler strains (FlyC31 strains) at BestGene Inc.

Currently available Perrimon strains (P{CaryP} strains) at BestGene Inc.

Other strains^*

updated 10/23/2024

Highlighted are popular/preferred strains