Co-expression of multiple transgenes in mouse CNS

A comparison of strategies

Joanna L. Jankowsky, Hilda H. Slunt, Tamara Ratovitski, Nancy A. Jenkins, Neal G. Copeland, David R. Borchelt

Research output: Contribution to journalArticle

Abstract

The introduction of two transgenes into one animal is increasingly common as transgenic experiments become more sophisticated. In this study we examine two strategies for creating double transgenic founders from a single microinjection. In the first approach, two constructs, each with its own promoter element, were coinjected into the pronucleus. In the second approach, both transgenes were cloned into one vector, separated by an internal ribosomal entry site (IRES), and placed under control of a single promoter. Both strategies save time and increase the percentage of double transgenic offspring over the standard method of mating single transgenic lines. However, despite high transgene copy numbers, the bicistronic lines did not show robust expression of either protein. Copy number and protein expression correlated much better in the coinjected lines, with expression levels in one line approaching that observed in some of our best single transgenic controls. Thus we recommend coinjection of individual plasmids for the generation of multiply transgenic founders.

Original languageEnglish (US)
Pages (from-to)157-165
Number of pages9
JournalBiomolecular Engineering
Volume17
Issue number6
DOIs
StatePublished - 2001

Fingerprint

Transgenes
Proteins
Animals
Plasmids
Microinjections
Experiments

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Jankowsky, J. L., Slunt, H. H., Ratovitski, T., Jenkins, N. A., Copeland, N. G., & Borchelt, D. R. (2001). Co-expression of multiple transgenes in mouse CNS: A comparison of strategies. Biomolecular Engineering, 17(6), 157-165. https://doi.org/10.1016/S1389-0344(01)00067-3

Co-expression of multiple transgenes in mouse CNS : A comparison of strategies. / Jankowsky, Joanna L.; Slunt, Hilda H.; Ratovitski, Tamara; Jenkins, Nancy A.; Copeland, Neal G.; Borchelt, David R.

In: Biomolecular Engineering, Vol. 17, No. 6, 2001, p. 157-165.

Research output: Contribution to journalArticle

Jankowsky, JL, Slunt, HH, Ratovitski, T, Jenkins, NA, Copeland, NG & Borchelt, DR 2001, 'Co-expression of multiple transgenes in mouse CNS: A comparison of strategies', Biomolecular Engineering, vol. 17, no. 6, pp. 157-165. https://doi.org/10.1016/S1389-0344(01)00067-3
Jankowsky, Joanna L. ; Slunt, Hilda H. ; Ratovitski, Tamara ; Jenkins, Nancy A. ; Copeland, Neal G. ; Borchelt, David R. / Co-expression of multiple transgenes in mouse CNS : A comparison of strategies. In: Biomolecular Engineering. 2001 ; Vol. 17, No. 6. pp. 157-165.
@article{8a617cdc240a4bd69a96b2b5e27fff29,
title = "Co-expression of multiple transgenes in mouse CNS: A comparison of strategies",
abstract = "The introduction of two transgenes into one animal is increasingly common as transgenic experiments become more sophisticated. In this study we examine two strategies for creating double transgenic founders from a single microinjection. In the first approach, two constructs, each with its own promoter element, were coinjected into the pronucleus. In the second approach, both transgenes were cloned into one vector, separated by an internal ribosomal entry site (IRES), and placed under control of a single promoter. Both strategies save time and increase the percentage of double transgenic offspring over the standard method of mating single transgenic lines. However, despite high transgene copy numbers, the bicistronic lines did not show robust expression of either protein. Copy number and protein expression correlated much better in the coinjected lines, with expression levels in one line approaching that observed in some of our best single transgenic controls. Thus we recommend coinjection of individual plasmids for the generation of multiply transgenic founders.",
author = "Jankowsky, {Joanna L.} and Slunt, {Hilda H.} and Tamara Ratovitski and Jenkins, {Nancy A.} and Copeland, {Neal G.} and Borchelt, {David R.}",
year = "2001",
doi = "10.1016/S1389-0344(01)00067-3",
language = "English (US)",
volume = "17",
pages = "157--165",
journal = "New Biotechnology",
issn = "1871-6784",
publisher = "Elsevier",
number = "6",

}

TY - JOUR

T1 - Co-expression of multiple transgenes in mouse CNS

T2 - A comparison of strategies

AU - Jankowsky, Joanna L.

AU - Slunt, Hilda H.

AU - Ratovitski, Tamara

AU - Jenkins, Nancy A.

AU - Copeland, Neal G.

AU - Borchelt, David R.

PY - 2001

Y1 - 2001

N2 - The introduction of two transgenes into one animal is increasingly common as transgenic experiments become more sophisticated. In this study we examine two strategies for creating double transgenic founders from a single microinjection. In the first approach, two constructs, each with its own promoter element, were coinjected into the pronucleus. In the second approach, both transgenes were cloned into one vector, separated by an internal ribosomal entry site (IRES), and placed under control of a single promoter. Both strategies save time and increase the percentage of double transgenic offspring over the standard method of mating single transgenic lines. However, despite high transgene copy numbers, the bicistronic lines did not show robust expression of either protein. Copy number and protein expression correlated much better in the coinjected lines, with expression levels in one line approaching that observed in some of our best single transgenic controls. Thus we recommend coinjection of individual plasmids for the generation of multiply transgenic founders.

AB - The introduction of two transgenes into one animal is increasingly common as transgenic experiments become more sophisticated. In this study we examine two strategies for creating double transgenic founders from a single microinjection. In the first approach, two constructs, each with its own promoter element, were coinjected into the pronucleus. In the second approach, both transgenes were cloned into one vector, separated by an internal ribosomal entry site (IRES), and placed under control of a single promoter. Both strategies save time and increase the percentage of double transgenic offspring over the standard method of mating single transgenic lines. However, despite high transgene copy numbers, the bicistronic lines did not show robust expression of either protein. Copy number and protein expression correlated much better in the coinjected lines, with expression levels in one line approaching that observed in some of our best single transgenic controls. Thus we recommend coinjection of individual plasmids for the generation of multiply transgenic founders.

UR - http://www.scopus.com/inward/record.url?scp=0035049961&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035049961&partnerID=8YFLogxK

U2 - 10.1016/S1389-0344(01)00067-3

DO - 10.1016/S1389-0344(01)00067-3

M3 - Article

VL - 17

SP - 157

EP - 165

JO - New Biotechnology

JF - New Biotechnology

SN - 1871-6784

IS - 6

ER -