Cytoplasmic expression systems triggered by mRNA yield increased gene expression in post-mitotic neurons

Research output: Contribution to journalArticle

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Cytoplasmic expression systems triggered by mRNA yield increased gene expression in post-mitotic neurons. / Farrow, PJ; Barrett, LB; Stevenson, M; Fisher, KD; Finn, J; Spice, Rachel; [No Value], [No Value]; Berry, Martin; Logan, Ann; Seymour, LW; Read, Martin.

In: Nucleic Acids Research, Vol. 34(11), No. e80, 28.06.2006, p. 1-12.

Research output: Contribution to journalArticle

Harvard

Farrow, PJ, Barrett, LB, Stevenson, M, Fisher, KD, Finn, J, Spice, R, [No Value], NV, Berry, M, Logan, A, Seymour, LW & Read, M 2006, 'Cytoplasmic expression systems triggered by mRNA yield increased gene expression in post-mitotic neurons', Nucleic Acids Research, vol. 34(11), no. e80, pp. 1-12. https://doi.org/10.1093/nar/gkl442

APA

Farrow, PJ., Barrett, LB., Stevenson, M., Fisher, KD., Finn, J., Spice, R., [No Value], N. V., Berry, M., Logan, A., Seymour, LW., & Read, M. (2006). Cytoplasmic expression systems triggered by mRNA yield increased gene expression in post-mitotic neurons. Nucleic Acids Research, 34(11)(e80), 1-12. https://doi.org/10.1093/nar/gkl442

Vancouver

Author

Farrow, PJ ; Barrett, LB ; Stevenson, M ; Fisher, KD ; Finn, J ; Spice, Rachel ; [No Value], [No Value] ; Berry, Martin ; Logan, Ann ; Seymour, LW ; Read, Martin. / Cytoplasmic expression systems triggered by mRNA yield increased gene expression in post-mitotic neurons. In: Nucleic Acids Research. 2006 ; Vol. 34(11), No. e80. pp. 1-12.

Bibtex

@article{eab3def56a4c447281cda43292e25055,
title = "Cytoplasmic expression systems triggered by mRNA yield increased gene expression in post-mitotic neurons",
abstract = "Non-viral vectors are promising vehicles for gene therapy but delivery of plasmid DNA to post-mitotic cells is challenging as nuclear entry is particularly inefficient. We have developed and evaluated a hybrid mRNA/DNA system designed to bypass the nuclear barrier to transfection and facilitate cytoplasmic gene expression. This system, based on co-delivery of mRNA(A64) encoding for T7 RNA polymerase (T7 RNAP) with a T7-driven plasmid, produced between 10- and 2200-fold higher gene expression in primary dorsal root ganglion neuronal (DRGN) cultures isolated from Sprague-Dawley rats compared to a cytomegalovirus (CMV)-driven plasmid, and 30-fold greater expression than the enhanced T7-based autogene plasmid pR011. Cell-free assays and in vitro transfections highlighted the versatility of this system with small quantities of T7 RNAP mRNA required to mediate expression at levels that were significantly greater than with the T7-driven plasmid alone or supplemented with T7 RNAP protein. We have also characterized a number of parameters, such as mRNA structure, intracellular stability and persistence of each nucleic acid component that represent important factors in determining the transfection efficiency of this hybrid expression system. The results from this study demonstrate that co-delivery of mRNA is a promising strategy to yield increased expression with plasmid DNA, and represents an important step towards improving the capability of non-viral vectors to mediate efficient gene transfer in cell types, such as in DRGN, where the nuclear membrane is a significant barrier to transfection.",
author = "PJ Farrow and LB Barrett and M Stevenson and KD Fisher and J Finn and Rachel Spice and {[No Value]}, {[No Value]} and Martin Berry and Ann Logan and LW Seymour and Martin Read",
year = "2006",
month = jun,
day = "28",
doi = "10.1093/nar/gkl442",
language = "English",
volume = "34(11)",
pages = "1--12",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "e80",

}

RIS

TY - JOUR

T1 - Cytoplasmic expression systems triggered by mRNA yield increased gene expression in post-mitotic neurons

AU - Farrow, PJ

AU - Barrett, LB

AU - Stevenson, M

AU - Fisher, KD

AU - Finn, J

AU - Spice, Rachel

AU - [No Value], [No Value]

AU - Berry, Martin

AU - Logan, Ann

AU - Seymour, LW

AU - Read, Martin

PY - 2006/6/28

Y1 - 2006/6/28

N2 - Non-viral vectors are promising vehicles for gene therapy but delivery of plasmid DNA to post-mitotic cells is challenging as nuclear entry is particularly inefficient. We have developed and evaluated a hybrid mRNA/DNA system designed to bypass the nuclear barrier to transfection and facilitate cytoplasmic gene expression. This system, based on co-delivery of mRNA(A64) encoding for T7 RNA polymerase (T7 RNAP) with a T7-driven plasmid, produced between 10- and 2200-fold higher gene expression in primary dorsal root ganglion neuronal (DRGN) cultures isolated from Sprague-Dawley rats compared to a cytomegalovirus (CMV)-driven plasmid, and 30-fold greater expression than the enhanced T7-based autogene plasmid pR011. Cell-free assays and in vitro transfections highlighted the versatility of this system with small quantities of T7 RNAP mRNA required to mediate expression at levels that were significantly greater than with the T7-driven plasmid alone or supplemented with T7 RNAP protein. We have also characterized a number of parameters, such as mRNA structure, intracellular stability and persistence of each nucleic acid component that represent important factors in determining the transfection efficiency of this hybrid expression system. The results from this study demonstrate that co-delivery of mRNA is a promising strategy to yield increased expression with plasmid DNA, and represents an important step towards improving the capability of non-viral vectors to mediate efficient gene transfer in cell types, such as in DRGN, where the nuclear membrane is a significant barrier to transfection.

AB - Non-viral vectors are promising vehicles for gene therapy but delivery of plasmid DNA to post-mitotic cells is challenging as nuclear entry is particularly inefficient. We have developed and evaluated a hybrid mRNA/DNA system designed to bypass the nuclear barrier to transfection and facilitate cytoplasmic gene expression. This system, based on co-delivery of mRNA(A64) encoding for T7 RNA polymerase (T7 RNAP) with a T7-driven plasmid, produced between 10- and 2200-fold higher gene expression in primary dorsal root ganglion neuronal (DRGN) cultures isolated from Sprague-Dawley rats compared to a cytomegalovirus (CMV)-driven plasmid, and 30-fold greater expression than the enhanced T7-based autogene plasmid pR011. Cell-free assays and in vitro transfections highlighted the versatility of this system with small quantities of T7 RNAP mRNA required to mediate expression at levels that were significantly greater than with the T7-driven plasmid alone or supplemented with T7 RNAP protein. We have also characterized a number of parameters, such as mRNA structure, intracellular stability and persistence of each nucleic acid component that represent important factors in determining the transfection efficiency of this hybrid expression system. The results from this study demonstrate that co-delivery of mRNA is a promising strategy to yield increased expression with plasmid DNA, and represents an important step towards improving the capability of non-viral vectors to mediate efficient gene transfer in cell types, such as in DRGN, where the nuclear membrane is a significant barrier to transfection.

UR - http://www.scopus.com/inward/record.url?scp=33746197965&partnerID=8YFLogxK

U2 - 10.1093/nar/gkl442

DO - 10.1093/nar/gkl442

M3 - Article

C2 - 16835307

VL - 34(11)

SP - 1

EP - 12

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - e80

ER -