DETECTION OF RIBONUCLEASE H-GENERATED MESSENGER-RNA FRAGMENTS IN HUMAN LEUKEMIA-CELLS FOLLOWING REVERSIBLE MEMBRANE PERMEABILIZATION IN THE PRESENCE OF ANTISENSE OLIGODEOXYNUCLEOTIDES

R V Giles; D G Spiller; D M Tidd

DETECTION OF RIBONUCLEASE H-GENERATED MESSENGER-RNA FRAGMENTS IN HUMAN LEUKEMIA-CELLS FOLLOWING REVERSIBLE MEMBRANE PERMEABILIZATION IN THE PRESENCE OF ANTISENSE OLIGODEOXYNUCLEOTIDES

R V Giles, D G Spiller, D M Tidd

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

Abstract

The involvement of ribonuclease H (RNase H) in antisense phenomena in intact cells has, to date, only been adequately demonstrated for microinjected Xenopus systems, The significance of RNase H for the antisense effects of oligodeoxynucleotides observed in human and other mammalian cell cultures has remained obscure, in part because of inadequate analytic methods. In this report we show that the ''reverse ligation-mediated PCR'' (RL-PCR) procedure permits amplification of RNA fragments produced by oligodeoxynucleotide-directed RNase H activity, We have used this procedure to demonstrate RNase H-dependent antisense effects in irreversibly permeabilized (dead) cells and reversibly permeabilized (live) cells.

Original language	English
Pages (from-to)	23-31
Number of pages	9
Journal	Antisense Research and Development
Volume	5
Issue number	1
Publication status	Published - 1995

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title = "DETECTION OF RIBONUCLEASE H-GENERATED MESSENGER-RNA FRAGMENTS IN HUMAN LEUKEMIA-CELLS FOLLOWING REVERSIBLE MEMBRANE PERMEABILIZATION IN THE PRESENCE OF ANTISENSE OLIGODEOXYNUCLEOTIDES",

abstract = "The involvement of ribonuclease H (RNase H) in antisense phenomena in intact cells has, to date, only been adequately demonstrated for microinjected Xenopus systems, The significance of RNase H for the antisense effects of oligodeoxynucleotides observed in human and other mammalian cell cultures has remained obscure, in part because of inadequate analytic methods. In this report we show that the ''reverse ligation-mediated PCR'' (RL-PCR) procedure permits amplification of RNA fragments produced by oligodeoxynucleotide-directed RNase H activity, We have used this procedure to demonstrate RNase H-dependent antisense effects in irreversibly permeabilized (dead) cells and reversibly permeabilized (live) cells.",

author = "Giles, {R V} and Spiller, {D G} and Tidd, {D M}",

year = "1995",

language = "English",

volume = "5",

pages = "23--31",

journal = "Antisense Research and Development",

issn = "1050-5261",

publisher = "Mary Ann Liebert Incorporated",

number = "1",

}

TY - JOUR

T1 - DETECTION OF RIBONUCLEASE H-GENERATED MESSENGER-RNA FRAGMENTS IN HUMAN LEUKEMIA-CELLS FOLLOWING REVERSIBLE MEMBRANE PERMEABILIZATION IN THE PRESENCE OF ANTISENSE OLIGODEOXYNUCLEOTIDES

AU - Giles, R V

AU - Spiller, D G

AU - Tidd, D M

PY - 1995

Y1 - 1995

N2 - The involvement of ribonuclease H (RNase H) in antisense phenomena in intact cells has, to date, only been adequately demonstrated for microinjected Xenopus systems, The significance of RNase H for the antisense effects of oligodeoxynucleotides observed in human and other mammalian cell cultures has remained obscure, in part because of inadequate analytic methods. In this report we show that the ''reverse ligation-mediated PCR'' (RL-PCR) procedure permits amplification of RNA fragments produced by oligodeoxynucleotide-directed RNase H activity, We have used this procedure to demonstrate RNase H-dependent antisense effects in irreversibly permeabilized (dead) cells and reversibly permeabilized (live) cells.

AB - The involvement of ribonuclease H (RNase H) in antisense phenomena in intact cells has, to date, only been adequately demonstrated for microinjected Xenopus systems, The significance of RNase H for the antisense effects of oligodeoxynucleotides observed in human and other mammalian cell cultures has remained obscure, in part because of inadequate analytic methods. In this report we show that the ''reverse ligation-mediated PCR'' (RL-PCR) procedure permits amplification of RNA fragments produced by oligodeoxynucleotide-directed RNase H activity, We have used this procedure to demonstrate RNase H-dependent antisense effects in irreversibly permeabilized (dead) cells and reversibly permeabilized (live) cells.

M3 - Article

SN - 1050-5261

VL - 5

SP - 23

EP - 31

JO - Antisense Research and Development

JF - Antisense Research and Development

IS - 1

ER -

DETECTION OF RIBONUCLEASE H-GENERATED MESSENGER-RNA FRAGMENTS IN HUMAN LEUKEMIA-CELLS FOLLOWING REVERSIBLE MEMBRANE PERMEABILIZATION IN THE PRESENCE OF ANTISENSE OLIGODEOXYNUCLEOTIDES

Abstract

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