ERRATUM TO

The GAL4 System: A Versatile System for the Manipulation

and Analysis of Gene Expression
Elizabeth E. Caygill and Andrea H. Brand

Christian Dahmann (ed.), Drosophila: Methods and Protocols, Methods in Molecular Biology, vol. 1478,
DOI 10.1007/978-1-4939-6371-3_2, © Springer Science+Business Media New York 2017

DOI 10.1007/978-1-4939-6371-3_22

In Chapter 02 titled “The GAL4 System: A Versatile System for the Manipulation and
Analysis of Gene Expression”, there was an error with the formatting in figures 3 and 4.
This has been changed to as follows:

The updated online version of the original chapter can be found at

http://dx.doi.org/10.1007/978-1-4939-6371-3_2

Christian Dahmann (ed.), Drosophila: Methods and Protocols, Methods in Molecular Biology, vol. 1478,
DOI 10.1007/978-1-4939-6371-3_22, © Springer Science+Business Media New York 2017

E1
 A. FLP-out GAL4:

Initial State:

Act5C yellow GAL4

FRT termination FRT
signals
FLP
Heat shock
promoter
marker
UAS

Heat Shock (37oC):

Act5C GAL4

FLP
Act5C GAL4
Heat shock
promoter

marker marker
UAS UAS

B. MARCM:

Initial State:
tub Gal80
FLP
Heat shock
promoter mutation

tub GAL4
marker

Heat Shock (37oC):

tub Gal80
tub Gal80

tub Gal80 tub Gal80

FLP
Heat shock
promoter

mutation mutation

tub GAL4 mutation mutation

marker
marker

Fig. 3 Clonal analysis. (a) Expression of UAS constructs in clones can be achieved using FLP-out GAL4. Heat
shock at 37 °C induces expression of a heat shock-inducible FLP recombinase. The recombinase acts on the FLP-out
cassette catalyzing recombination between direct FRT repeats. Recombination removes the yellow marker and the tran-
scription termination signals, allowing the expression of GAL4 under control of the Actin promoter. (b) Positively marked
mutant clones can be made using MARCM. Heat shock at 37 °C induces expression of FLP recombinase that catalyzes
recombination between FRTs on homologous chromosomes carrying either the mutation of interest or a tubGAL80 construct.
Segregation of the GAL80 from the mutation into different daughter cells relieves repression of a GAL4 inducible marker in
the mutant cells, labeling the mutant clone
 Erratum E3

A. UAS-IR
target mRNA
AA

A
AA
Enhancer GAL4
UAS Inverted Repeat Sequence

mRNA degradation
AA

A
AA
B. UAS-shmiR
target mRNA
AA

A
AA
Enhancer GAL4
UAS shmiR Sequence

AA mRNA degradation

A
AA
C. UAS-cas9

Enhancer GAL4 cas9

UAS

target DNA

U6:3 double strand break

gRNA

homologous recombination
or
non-homologous end joining

Fig. 4 Repressing genes using the GAL4 system. (a) RNAi-based gene knockdown can be achieved by expressing inverted
repeats complementary to the gene of interest. These repeats will fold into a long double-stranded RNA that will be pro-
cessed to produce multiple siRNAs against the target gene. (b) More specific knockdown can be achieved using UAS-
shmiR constructs. A single targeting siRNA is cloned into the miR-1 backbone. Processing by cellular machinery produces
only that siRNA. (c) Tissue-specific gene knock-outs can be achieved using a GAL4-dependent version of Cas9 and the
CRISPR/Cas9 gene targeting. A ubiquitously expressed chimeric gRNA targeting the gene of interest is expressed in con-
junction with UAS-cas9 resulting in biallelic gene targeting where Cas9 is driven