drivers listed below drive predominantly in one or other of these
domains. Also listed is pnr-GAL4 which express strongly in both
the PE cells, near the stalk region, and the underlying disc proper
(DP) epithelial cells that will give rise to the central notum region.
Although each of the peripodial GAL4 drivers can elicit eversion
phenotypes, we find Ubx-GAL4 is the most effective at producing
clear eversion-failure phenotypes such as internalized wings. The
pnr-GAL4 driver can also elicit eversion failure, though more commonly it results in disruption of the later epithelial migration and
fusion events, and produces thoracic clefts [23]. Also listed are
stocks carrying GAL80
ts and UAS-Dicer-2 constructs, which can
be combined with the GAL4 constructs to achieve more effective
driver stocks.
2.1.2 RNAi Libraries
Three public stock collections are available for GAL4-UAS RNAi
knockdown (Table 2). These have been thoroughly described in a
recent methods paper [2] and so will be only briefly covered here.
We have achieved strong eversion phenotypes with UAS lines from
each of these libraries.
Table 2
UAS RNAi libraries
Library Source
Comment
URL
VDRC
[23]
The Vienna Drosophila
Resource Centre, Vienna,
Austria
12,671 stocks covering 91% of proteincoding genes.
Three libraries are available: GD library:
Long hairpin dsRNA constructs,
random P-element insertions
KK library: Long hairpin dsRNA
constructs, attP insertions on
chromosome II [insertions are
potentially at two sites: 40D3 and
30B3] [29, 30]
shRNA library: Short hairpin RNA
constructs inserted at {attP}40 at 25C6
http://
stockcenter.
vdrc.at
NIGFLY
The National Institute for
Genetics, Mishima, Japan
11,910 stocks covering 45% of proteincoding genes.
Long hairpin dsRNA constructs, random
P-element insertions
http://www.
shigen.nig.ac.
jp/fly/nigfly/
TRiP
[11, 31,
32]
The Transgenic RNAi Project
at the Harvard Medical
School, Boston, USA
11,454 stocks covering 70% of proteincoding genes.
Long hairpin dsRNA constructs
(VALIUM1 and VALIUM10) OR short
hairpin constructs (VALIUM
20, VALIUM21, and VALIUM22).
Inserts on chromosomes II ({attP}40 at
25C6) and III ({attP}2 at 68A4)
http://www.
flyrnai.org/
TRiPHOME.html
Using Drosophila Wing Eversion to Study EMT
119
domains. Also listed is pnr-GAL4 which express strongly in both
the PE cells, near the stalk region, and the underlying disc proper
(DP) epithelial cells that will give rise to the central notum region.
Although each of the peripodial GAL4 drivers can elicit eversion
phenotypes, we find Ubx-GAL4 is the most effective at producing
clear eversion-failure phenotypes such as internalized wings. The
pnr-GAL4 driver can also elicit eversion failure, though more commonly it results in disruption of the later epithelial migration and
fusion events, and produces thoracic clefts [23]. Also listed are
stocks carrying GAL80
ts and UAS-Dicer-2 constructs, which can
be combined with the GAL4 constructs to achieve more effective
driver stocks.
2.1.2 RNAi Libraries
Three public stock collections are available for GAL4-UAS RNAi
knockdown (Table 2). These have been thoroughly described in a
recent methods paper [2] and so will be only briefly covered here.
We have achieved strong eversion phenotypes with UAS lines from
each of these libraries.
Table 2
UAS RNAi libraries
Library Source
Comment
URL
VDRC
[23]
The Vienna Drosophila
Resource Centre, Vienna,
Austria
12,671 stocks covering 91% of proteincoding genes.
Three libraries are available: GD library:
Long hairpin dsRNA constructs,
random P-element insertions
KK library: Long hairpin dsRNA
constructs, attP insertions on
chromosome II [insertions are
potentially at two sites: 40D3 and
30B3] [29, 30]
shRNA library: Short hairpin RNA
constructs inserted at {attP}40 at 25C6
http://
stockcenter.
vdrc.at
NIGFLY
The National Institute for
Genetics, Mishima, Japan
11,910 stocks covering 45% of proteincoding genes.
Long hairpin dsRNA constructs, random
P-element insertions
http://www.
shigen.nig.ac.
jp/fly/nigfly/
TRiP
[11, 31,
32]
The Transgenic RNAi Project
at the Harvard Medical
School, Boston, USA
11,454 stocks covering 70% of proteincoding genes.
Long hairpin dsRNA constructs
(VALIUM1 and VALIUM10) OR short
hairpin constructs (VALIUM
20, VALIUM21, and VALIUM22).
Inserts on chromosomes II ({attP}40 at
25C6) and III ({attP}2 at 68A4)
http://www.
flyrnai.org/
TRiPHOME.html
Using Drosophila Wing Eversion to Study EMT
119