A LexAop > UAS > QUAS trimeric plasmid to generate inducible and interconvertible Drosophila overexpression transgenes

Franz Wendler; Sangbin Park; Claire Hill; Alessia Galasso; Kathleen R. Chang; Iman Awan; Yulia Sudarikova; Mar Bustamante-Sequeiros; Sichen Liu; Ethan Y.H. Sung; Gabrielle Aisa-Bonoko; Seung K. Kim; Luis A. Baena-Lopez

doi:10.1038/s41598-022-07852-7

A LexAop > UAS > QUAS trimeric plasmid to generate inducible and interconvertible Drosophila overexpression transgenes

Franz Wendler, Sangbin Park, Claire Hill, Alessia Galasso, Kathleen R. Chang, Iman Awan, Yulia Sudarikova, Mar Bustamante-Sequeiros, Sichen Liu, Ethan Y.H. Sung, Gabrielle Aisa-Bonoko, Seung K. Kim^*, Luis A. Baena-Lopez^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

17 Downloads (Pure)

Abstract

The existence of three independent binary systems for conditional gene expression (Gal4/UAS; LexA/LexAop; QF/QUAS) has greatly expanded versatile genetic analyses in the Drosophila melanogaster; however, the experimental application of these tools is limited by the need to generate multiple collections of noninterchangeable transgenic fly strains for each inducible gene expression system. To address this practical limitation, we developed a modular vector that contains the regulatory elements from all three binary systems, enabling Gal4-, LexA- or QF-dependent expression of transgenes. Our methods also incorporate DNA elements that facilitate independent site-specific recombination and elimination of regulatory UAS, LexAop or QUAS modules with spatial and temporal control, thus offering unprecedented possibilities and logistical advantages for in vivo genetic modulation and efficient interconversion of overexpression transgenic fly lines.

Original language	English
Article number	3835
Number of pages	10
Journal	Scientific Reports
Volume	12
Early online date	09 Mar 2022
DOIs	https://doi.org/10.1038/s41598-022-07852-7
Publication status	Published - Dec 2022
Externally published	Yes

Bibliographical note

Funding Information:
We are grateful to the following investigators for providing flies and reagents: Drs. I. Salecker (for kindly providing all the reagents and flies used in the study related to mFlp5 and the hs-Cre line on the second chromosome), K. Basler (for providing the βTub85D-Flippase and dpp-LexA strains), Bloomington Stock Center (fly strains), Kyoto Stock Center (fly strains) and DGRC (wild-type cDNAs). We thank Dr. G. Struhl for sharing information regarding the βTub85D promoter and Genewiz and Bestgene for DNA synthesis and generating the transgenic flies, respectively. We are also grateful for support from the Stanford Bing Overseas Program at Oxford, directed by Dr. S. Sodywola. We also thank the caspaselab members ( https://www.caspaselab.com ) at Oxford for critical reading of the manuscript and valuable suggestions. This work was supported by Cancer Research UK C49979/A17516, the Edward Penley Abraham Research Funds (RF290 and RF286 (19)), and the John Fell Fund from the University of Oxford 162/001. L.A.B-L. is a CRUK Career Development Fellow (C49979/A17516) and an Oriel College Hayward Fellow. C.H. is a PhD student in the BBSRC-funded Oxford Interdisciplinary Bioscience Doctoral Training Programme (BB/M011224/1). A.G. and F. W are postdoctoral fellows of CRUK (C49979/A17516). K.R.C. was supported by a Vice Provost Undergraduate Education (VPUE) fellowship from Stanford University. Work here was supported by a VPUE Course Development Grant and by NIH awards (R01 DK107507; R01 DK108817; U01 DK123743 to S.K.K.), the JDRF Northern California Center of Excellence, and by NIH grant P30 DK116074 (S.K.K.) for the Stanford Diabetes Research Center. WT

Funding Information:
We are grateful to the following investigators for providing flies and reagents: Drs. I. Salecker (for kindly providing all the reagents and flies used in the study related to mFlp5 and the hs-Cre line on the second chromosome), K. Basler (for providing the ?Tub85D -FlippaseWT and dpp -LexA strains), Bloomington Stock Center (fly strains), Kyoto Stock Center (fly strains) and DGRC (wild-type cDNAs). We thank Dr. G. Struhl for sharing information regarding the ?Tub85D promoter and Genewiz and Bestgene for DNA synthesis and generating the transgenic flies, respectively. We are also grateful for support from the Stanford Bing Overseas Program at Oxford, directed by Dr. S. Sodywola. We also thank the caspaselab members (https://www.caspaselab.com) at Oxford for critical reading of the manuscript and valuable suggestions. This work was supported by Cancer Research UK C49979/A17516, the Edward Penley Abraham Research Funds (RF290 and RF286 (19)), and the John Fell Fund from the University of Oxford 162/001. L.A.B-L. is a CRUK Career Development Fellow (C49979/A17516) and an Oriel College Hayward Fellow. C.H. is a PhD student in the BBSRC-funded Oxford Interdisciplinary Bioscience Doctoral Training Programme (BB/M011224/1). A.G. and F. W are postdoctoral fellows of CRUK (C49979/A17516). K.R.C. was supported by a Vice Provost Undergraduate Education (VPUE) fellowship from Stanford University. Work here was supported by a VPUE Course Development Grant and by NIH awards (R01 DK107507; R01 DK108817; U01 DK123743 to S.K.K.), the JDRF Northern California Center of Excellence, and by NIH grant P30 DK116074 (S.K.K.) for the Stanford Diabetes Research Center.

Publisher Copyright:
© 2022, The Author(s).

ASJC Scopus subject areas

General

Access to Document

10.1038/s41598-022-07852-7Licence: CC BY

A LexAop>UAS>QUAS trimeric plasmid to generate inducible and interconvertible Drosophila overexpression transgenes
© 2023 The Authors. This is an open access article published under a Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
Final published version, 2.59 MBLicence: CC BY

Cite this

Wendler, F., Park, S., Hill, C., Galasso, A., Chang, K. R., Awan, I., Sudarikova, Y., Bustamante-Sequeiros, M., Liu, S., Sung, E. Y. H., Aisa-Bonoko, G., Kim, S. K., & Baena-Lopez, L. A. (2022). A LexAop > UAS > QUAS trimeric plasmid to generate inducible and interconvertible Drosophila overexpression transgenes. Scientific Reports, 12, Article 3835. https://doi.org/10.1038/s41598-022-07852-7

@article{1cf3314ff8994062bdaf13ef87426da2,

title = "A LexAop > UAS > QUAS trimeric plasmid to generate inducible and interconvertible Drosophila overexpression transgenes",

abstract = "The existence of three independent binary systems for conditional gene expression (Gal4/UAS; LexA/LexAop; QF/QUAS) has greatly expanded versatile genetic analyses in the Drosophila melanogaster; however, the experimental application of these tools is limited by the need to generate multiple collections of noninterchangeable transgenic fly strains for each inducible gene expression system. To address this practical limitation, we developed a modular vector that contains the regulatory elements from all three binary systems, enabling Gal4-, LexA- or QF-dependent expression of transgenes. Our methods also incorporate DNA elements that facilitate independent site-specific recombination and elimination of regulatory UAS, LexAop or QUAS modules with spatial and temporal control, thus offering unprecedented possibilities and logistical advantages for in vivo genetic modulation and efficient interconversion of overexpression transgenic fly lines.",

author = "Franz Wendler and Sangbin Park and Claire Hill and Alessia Galasso and Chang, {Kathleen R.} and Iman Awan and Yulia Sudarikova and Mar Bustamante-Sequeiros and Sichen Liu and Sung, {Ethan Y.H.} and Gabrielle Aisa-Bonoko and Kim, {Seung K.} and Baena-Lopez, {Luis A.}",

note = "Funding Information: We are grateful to the following investigators for providing flies and reagents: Drs. I. Salecker (for kindly providing all the reagents and flies used in the study related to mFlp5 and the hs-Cre line on the second chromosome), K. Basler (for providing the βTub85D-Flippase and dpp-LexA strains), Bloomington Stock Center (fly strains), Kyoto Stock Center (fly strains) and DGRC (wild-type cDNAs). We thank Dr. G. Struhl for sharing information regarding the βTub85D promoter and Genewiz and Bestgene for DNA synthesis and generating the transgenic flies, respectively. We are also grateful for support from the Stanford Bing Overseas Program at Oxford, directed by Dr. S. Sodywola. We also thank the caspaselab members ( https://www.caspaselab.com ) at Oxford for critical reading of the manuscript and valuable suggestions. This work was supported by Cancer Research UK C49979/A17516, the Edward Penley Abraham Research Funds (RF290 and RF286 (19)), and the John Fell Fund from the University of Oxford 162/001. L.A.B-L. is a CRUK Career Development Fellow (C49979/A17516) and an Oriel College Hayward Fellow. C.H. is a PhD student in the BBSRC-funded Oxford Interdisciplinary Bioscience Doctoral Training Programme (BB/M011224/1). A.G. and F. W are postdoctoral fellows of CRUK (C49979/A17516). K.R.C. was supported by a Vice Provost Undergraduate Education (VPUE) fellowship from Stanford University. Work here was supported by a VPUE Course Development Grant and by NIH awards (R01 DK107507; R01 DK108817; U01 DK123743 to S.K.K.), the JDRF Northern California Center of Excellence, and by NIH grant P30 DK116074 (S.K.K.) for the Stanford Diabetes Research Center. WT Funding Information: We are grateful to the following investigators for providing flies and reagents: Drs. I. Salecker (for kindly providing all the reagents and flies used in the study related to mFlp5 and the hs-Cre line on the second chromosome), K. Basler (for providing the ?Tub85D -FlippaseWT and dpp -LexA strains), Bloomington Stock Center (fly strains), Kyoto Stock Center (fly strains) and DGRC (wild-type cDNAs). We thank Dr. G. Struhl for sharing information regarding the ?Tub85D promoter and Genewiz and Bestgene for DNA synthesis and generating the transgenic flies, respectively. We are also grateful for support from the Stanford Bing Overseas Program at Oxford, directed by Dr. S. Sodywola. We also thank the caspaselab members (https://www.caspaselab.com) at Oxford for critical reading of the manuscript and valuable suggestions. This work was supported by Cancer Research UK C49979/A17516, the Edward Penley Abraham Research Funds (RF290 and RF286 (19)), and the John Fell Fund from the University of Oxford 162/001. L.A.B-L. is a CRUK Career Development Fellow (C49979/A17516) and an Oriel College Hayward Fellow. C.H. is a PhD student in the BBSRC-funded Oxford Interdisciplinary Bioscience Doctoral Training Programme (BB/M011224/1). A.G. and F. W are postdoctoral fellows of CRUK (C49979/A17516). K.R.C. was supported by a Vice Provost Undergraduate Education (VPUE) fellowship from Stanford University. Work here was supported by a VPUE Course Development Grant and by NIH awards (R01 DK107507; R01 DK108817; U01 DK123743 to S.K.K.), the JDRF Northern California Center of Excellence, and by NIH grant P30 DK116074 (S.K.K.) for the Stanford Diabetes Research Center. Publisher Copyright: {\textcopyright} 2022, The Author(s).",

year = "2022",

month = dec,

doi = "10.1038/s41598-022-07852-7",

language = "English",

volume = "12",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - A LexAop > UAS > QUAS trimeric plasmid to generate inducible and interconvertible Drosophila overexpression transgenes

AU - Wendler, Franz

AU - Park, Sangbin

AU - Hill, Claire

AU - Galasso, Alessia

AU - Chang, Kathleen R.

AU - Awan, Iman

AU - Sudarikova, Yulia

AU - Bustamante-Sequeiros, Mar

AU - Liu, Sichen

AU - Sung, Ethan Y.H.

AU - Aisa-Bonoko, Gabrielle

AU - Kim, Seung K.

AU - Baena-Lopez, Luis A.

N1 - Funding Information: We are grateful to the following investigators for providing flies and reagents: Drs. I. Salecker (for kindly providing all the reagents and flies used in the study related to mFlp5 and the hs-Cre line on the second chromosome), K. Basler (for providing the βTub85D-Flippase and dpp-LexA strains), Bloomington Stock Center (fly strains), Kyoto Stock Center (fly strains) and DGRC (wild-type cDNAs). We thank Dr. G. Struhl for sharing information regarding the βTub85D promoter and Genewiz and Bestgene for DNA synthesis and generating the transgenic flies, respectively. We are also grateful for support from the Stanford Bing Overseas Program at Oxford, directed by Dr. S. Sodywola. We also thank the caspaselab members ( https://www.caspaselab.com ) at Oxford for critical reading of the manuscript and valuable suggestions. This work was supported by Cancer Research UK C49979/A17516, the Edward Penley Abraham Research Funds (RF290 and RF286 (19)), and the John Fell Fund from the University of Oxford 162/001. L.A.B-L. is a CRUK Career Development Fellow (C49979/A17516) and an Oriel College Hayward Fellow. C.H. is a PhD student in the BBSRC-funded Oxford Interdisciplinary Bioscience Doctoral Training Programme (BB/M011224/1). A.G. and F. W are postdoctoral fellows of CRUK (C49979/A17516). K.R.C. was supported by a Vice Provost Undergraduate Education (VPUE) fellowship from Stanford University. Work here was supported by a VPUE Course Development Grant and by NIH awards (R01 DK107507; R01 DK108817; U01 DK123743 to S.K.K.), the JDRF Northern California Center of Excellence, and by NIH grant P30 DK116074 (S.K.K.) for the Stanford Diabetes Research Center. WT Funding Information: We are grateful to the following investigators for providing flies and reagents: Drs. I. Salecker (for kindly providing all the reagents and flies used in the study related to mFlp5 and the hs-Cre line on the second chromosome), K. Basler (for providing the ?Tub85D -FlippaseWT and dpp -LexA strains), Bloomington Stock Center (fly strains), Kyoto Stock Center (fly strains) and DGRC (wild-type cDNAs). We thank Dr. G. Struhl for sharing information regarding the ?Tub85D promoter and Genewiz and Bestgene for DNA synthesis and generating the transgenic flies, respectively. We are also grateful for support from the Stanford Bing Overseas Program at Oxford, directed by Dr. S. Sodywola. We also thank the caspaselab members (https://www.caspaselab.com) at Oxford for critical reading of the manuscript and valuable suggestions. This work was supported by Cancer Research UK C49979/A17516, the Edward Penley Abraham Research Funds (RF290 and RF286 (19)), and the John Fell Fund from the University of Oxford 162/001. L.A.B-L. is a CRUK Career Development Fellow (C49979/A17516) and an Oriel College Hayward Fellow. C.H. is a PhD student in the BBSRC-funded Oxford Interdisciplinary Bioscience Doctoral Training Programme (BB/M011224/1). A.G. and F. W are postdoctoral fellows of CRUK (C49979/A17516). K.R.C. was supported by a Vice Provost Undergraduate Education (VPUE) fellowship from Stanford University. Work here was supported by a VPUE Course Development Grant and by NIH awards (R01 DK107507; R01 DK108817; U01 DK123743 to S.K.K.), the JDRF Northern California Center of Excellence, and by NIH grant P30 DK116074 (S.K.K.) for the Stanford Diabetes Research Center. Publisher Copyright: © 2022, The Author(s).

PY - 2022/12

Y1 - 2022/12

N2 - The existence of three independent binary systems for conditional gene expression (Gal4/UAS; LexA/LexAop; QF/QUAS) has greatly expanded versatile genetic analyses in the Drosophila melanogaster; however, the experimental application of these tools is limited by the need to generate multiple collections of noninterchangeable transgenic fly strains for each inducible gene expression system. To address this practical limitation, we developed a modular vector that contains the regulatory elements from all three binary systems, enabling Gal4-, LexA- or QF-dependent expression of transgenes. Our methods also incorporate DNA elements that facilitate independent site-specific recombination and elimination of regulatory UAS, LexAop or QUAS modules with spatial and temporal control, thus offering unprecedented possibilities and logistical advantages for in vivo genetic modulation and efficient interconversion of overexpression transgenic fly lines.

AB - The existence of three independent binary systems for conditional gene expression (Gal4/UAS; LexA/LexAop; QF/QUAS) has greatly expanded versatile genetic analyses in the Drosophila melanogaster; however, the experimental application of these tools is limited by the need to generate multiple collections of noninterchangeable transgenic fly strains for each inducible gene expression system. To address this practical limitation, we developed a modular vector that contains the regulatory elements from all three binary systems, enabling Gal4-, LexA- or QF-dependent expression of transgenes. Our methods also incorporate DNA elements that facilitate independent site-specific recombination and elimination of regulatory UAS, LexAop or QUAS modules with spatial and temporal control, thus offering unprecedented possibilities and logistical advantages for in vivo genetic modulation and efficient interconversion of overexpression transgenic fly lines.

U2 - 10.1038/s41598-022-07852-7

DO - 10.1038/s41598-022-07852-7

M3 - Article

C2 - 35264662

AN - SCOPUS:85126080970

SN - 2045-2322

VL - 12

JO - Scientific Reports

JF - Scientific Reports

M1 - 3835

ER -