Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon

Ann E. Davidson; Darius Balciunas; Deanna Mohn; Jennifer Shaffer; Spencer Hermanson; Sridhar Sivasubbu; M. Pat Cliff; Perry B. Hackett; Stephen C. Ekker

doi:10.1016/j.ydbio.2003.07.013

Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon

Ann E. Davidson, Darius Balciunas, Deanna Mohn, Jennifer Shaffer, Spencer Hermanson, Sridhar Sivasubbu, M. Pat Cliff, Perry B. Hackett, Stephen C. Ekker

Genetics, Cell Biology and Development (CBS)

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

202 Scopus citations

Abstract

We used the Tc1/mariner family transposable element Sleeping Beauty (SB) for transgenesis and long-term expression studies in the zebrafish (Danio rerio), a popular organism for clinical disease, vertebrate patterning, and cell biology applications. SB transposase enhanced the transgenesis and expression rate sixfold (from 5 to 31%) and more than doubled the total number of tagged chromosomes over standard, plasmid injection-based transgenesis methods. Molecular analysis of these loci demonstrated a precise integration of these elements into recipient chromosomes with genetic footprints diagnostic of transposition. GFP expression from transposase-mediated integrants was Mendelian through the eighth generation. A blue-shifted GFP variant (BFP) and a red fluorescent protein (DsRed) were also useful transgenesis markers, indicating that multiple reporters are practical for use with SB in zebrafish. We showed that SB is suitable for tissue-specific transgene applications using an abbreviated gamma-crystallin GFP cassette. Finally, we describe a general utility transposon vector for chromosomal engineering and molecular genetics experiments in zebrafish. Together, these data indicate that SB is an efficient tool for transgenesis and expression in zebrafish, and that the transposon will be useful for gene expression in cell biology applications as well as an insertional mutagen for gene discovery during development.

Original language	English (US)
Pages (from-to)	191-202
Number of pages	12
Journal	Developmental Biology
Volume	263
Issue number	2
DOIs	https://doi.org/10.1016/j.ydbio.2003.07.013
State	Published - Nov 15 2003

Bibliographical note

Funding Information:
We thank David Largaespada, Scott McIvor, Adam Dupuy, and Karl Clark for helpful discussions throughout the course of this work. For technical assistance, we give special thanks to Erin Walsh, Cheryl Saunders, Ken Finley, Daiva Urneziene, and Rachel Dries. We also thank Eleanor Chen and David Largaespada for comments on the manuscript. This work has been supported by the Arnold and Mabel Beckman Foundation and NIH Grants DA14546 and RRO6625.

Keywords

Insertional mutagenesis
Sleeping Beauty
Transgene expression
Transposon
Zebrafish

Access

10.1016/j.ydbio.2003.07.013

OpenUrl availability

Full text

Cite this

@article{e497bcb3890642aeb4978e1d61bacd0b,

title = "Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon",

abstract = "We used the Tc1/mariner family transposable element Sleeping Beauty (SB) for transgenesis and long-term expression studies in the zebrafish (Danio rerio), a popular organism for clinical disease, vertebrate patterning, and cell biology applications. SB transposase enhanced the transgenesis and expression rate sixfold (from 5 to 31%) and more than doubled the total number of tagged chromosomes over standard, plasmid injection-based transgenesis methods. Molecular analysis of these loci demonstrated a precise integration of these elements into recipient chromosomes with genetic footprints diagnostic of transposition. GFP expression from transposase-mediated integrants was Mendelian through the eighth generation. A blue-shifted GFP variant (BFP) and a red fluorescent protein (DsRed) were also useful transgenesis markers, indicating that multiple reporters are practical for use with SB in zebrafish. We showed that SB is suitable for tissue-specific transgene applications using an abbreviated gamma-crystallin GFP cassette. Finally, we describe a general utility transposon vector for chromosomal engineering and molecular genetics experiments in zebrafish. Together, these data indicate that SB is an efficient tool for transgenesis and expression in zebrafish, and that the transposon will be useful for gene expression in cell biology applications as well as an insertional mutagen for gene discovery during development.",

keywords = "Insertional mutagenesis, Sleeping Beauty, Transgene expression, Transposon, Zebrafish",

author = "Davidson, {Ann E.} and Darius Balciunas and Deanna Mohn and Jennifer Shaffer and Spencer Hermanson and Sridhar Sivasubbu and Cliff, {M. Pat} and Hackett, {Perry B.} and Ekker, {Stephen C.}",

note = "Funding Information: We thank David Largaespada, Scott McIvor, Adam Dupuy, and Karl Clark for helpful discussions throughout the course of this work. For technical assistance, we give special thanks to Erin Walsh, Cheryl Saunders, Ken Finley, Daiva Urneziene, and Rachel Dries. We also thank Eleanor Chen and David Largaespada for comments on the manuscript. This work has been supported by the Arnold and Mabel Beckman Foundation and NIH Grants DA14546 and RRO6625.",

year = "2003",

month = nov,

day = "15",

doi = "10.1016/j.ydbio.2003.07.013",

language = "English (US)",

volume = "263",

pages = "191--202",

journal = "Developmental Biology",

issn = "0012-1606",

publisher = "Academic Press Inc.",

number = "2",

}

TY - JOUR

T1 - Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon

AU - Davidson, Ann E.

AU - Balciunas, Darius

AU - Mohn, Deanna

AU - Shaffer, Jennifer

AU - Hermanson, Spencer

AU - Sivasubbu, Sridhar

AU - Cliff, M. Pat

AU - Hackett, Perry B.

AU - Ekker, Stephen C.

N1 - Funding Information: We thank David Largaespada, Scott McIvor, Adam Dupuy, and Karl Clark for helpful discussions throughout the course of this work. For technical assistance, we give special thanks to Erin Walsh, Cheryl Saunders, Ken Finley, Daiva Urneziene, and Rachel Dries. We also thank Eleanor Chen and David Largaespada for comments on the manuscript. This work has been supported by the Arnold and Mabel Beckman Foundation and NIH Grants DA14546 and RRO6625.

PY - 2003/11/15

Y1 - 2003/11/15

N2 - We used the Tc1/mariner family transposable element Sleeping Beauty (SB) for transgenesis and long-term expression studies in the zebrafish (Danio rerio), a popular organism for clinical disease, vertebrate patterning, and cell biology applications. SB transposase enhanced the transgenesis and expression rate sixfold (from 5 to 31%) and more than doubled the total number of tagged chromosomes over standard, plasmid injection-based transgenesis methods. Molecular analysis of these loci demonstrated a precise integration of these elements into recipient chromosomes with genetic footprints diagnostic of transposition. GFP expression from transposase-mediated integrants was Mendelian through the eighth generation. A blue-shifted GFP variant (BFP) and a red fluorescent protein (DsRed) were also useful transgenesis markers, indicating that multiple reporters are practical for use with SB in zebrafish. We showed that SB is suitable for tissue-specific transgene applications using an abbreviated gamma-crystallin GFP cassette. Finally, we describe a general utility transposon vector for chromosomal engineering and molecular genetics experiments in zebrafish. Together, these data indicate that SB is an efficient tool for transgenesis and expression in zebrafish, and that the transposon will be useful for gene expression in cell biology applications as well as an insertional mutagen for gene discovery during development.

AB - We used the Tc1/mariner family transposable element Sleeping Beauty (SB) for transgenesis and long-term expression studies in the zebrafish (Danio rerio), a popular organism for clinical disease, vertebrate patterning, and cell biology applications. SB transposase enhanced the transgenesis and expression rate sixfold (from 5 to 31%) and more than doubled the total number of tagged chromosomes over standard, plasmid injection-based transgenesis methods. Molecular analysis of these loci demonstrated a precise integration of these elements into recipient chromosomes with genetic footprints diagnostic of transposition. GFP expression from transposase-mediated integrants was Mendelian through the eighth generation. A blue-shifted GFP variant (BFP) and a red fluorescent protein (DsRed) were also useful transgenesis markers, indicating that multiple reporters are practical for use with SB in zebrafish. We showed that SB is suitable for tissue-specific transgene applications using an abbreviated gamma-crystallin GFP cassette. Finally, we describe a general utility transposon vector for chromosomal engineering and molecular genetics experiments in zebrafish. Together, these data indicate that SB is an efficient tool for transgenesis and expression in zebrafish, and that the transposon will be useful for gene expression in cell biology applications as well as an insertional mutagen for gene discovery during development.

KW - Insertional mutagenesis

KW - Sleeping Beauty

KW - Transgene expression

KW - Transposon

KW - Zebrafish

UR - http://www.scopus.com/inward/record.url?scp=0242334000&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0242334000&partnerID=8YFLogxK

U2 - 10.1016/j.ydbio.2003.07.013

DO - 10.1016/j.ydbio.2003.07.013

M3 - Article

C2 - 14597195

AN - SCOPUS:0242334000

SN - 0012-1606

VL - 263

SP - 191

EP - 202

JO - Developmental Biology

JF - Developmental Biology

IS - 2

ER -

Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon

Abstract

Bibliographical note

Keywords

Access

OpenUrl availability

Other files and links

Fingerprint

Cite this