Gel shift assay: Demonstration of enhanced binding of oligo(δ)-L-ornithine-oligodeoxynucleotide conjugates to complementary DNA and RNA

Tianmin Zhu, Shahriar Pooyan, Ziping Wei, Michael J Leibowitz, Stanley Stein

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

An increase in melting temperature for DNA:DNA duplexes had been observed previously (Zhu et al. Antisense Res. Dev. 3:349-356, 1993) when an oligo(δ)ornithine moiety was covalently appended to a short oligodeoxynucleotide. We now report the analysis of duplex formation by electrophoretic gel shift analysis. In the particular example studied, an increase in Tm of 4°C was found to correspond to about a fivefold increase in binding constant. A similar enhancement by the appended cationic peptide was observed when the target strand was RNA. The use of a competitive assay format for avoidance of adsorptive loss at low concentrations (<10-7 M) of the oligonucleotide-oligo(δ)ornithine conjugate is presented.

Original languageEnglish (US)
Pages (from-to)69-74
Number of pages6
JournalAntisense and Nucleic Acid Drug Development
Volume6
Issue number1
StatePublished - Mar 1996
Externally publishedYes

Fingerprint

Complementary RNA
Ornithine
Oligodeoxyribonucleotides
Assays
Demonstrations
Complementary DNA
Gels
Nucleic Acid Denaturation
DNA
Electrophoretic Mobility Shift Assay
Oligonucleotides
Melting point
RNA
Peptides
Temperature

ASJC Scopus subject areas

  • Genetics
  • Pharmacology

Cite this

Gel shift assay : Demonstration of enhanced binding of oligo(δ)-L-ornithine-oligodeoxynucleotide conjugates to complementary DNA and RNA. / Zhu, Tianmin; Pooyan, Shahriar; Wei, Ziping; Leibowitz, Michael J; Stein, Stanley.

In: Antisense and Nucleic Acid Drug Development, Vol. 6, No. 1, 03.1996, p. 69-74.

Research output: Contribution to journalArticle

@article{5a32c2de1bfe49e2b6e6d563eb3724bc,
title = "Gel shift assay: Demonstration of enhanced binding of oligo(δ)-L-ornithine-oligodeoxynucleotide conjugates to complementary DNA and RNA",
abstract = "An increase in melting temperature for DNA:DNA duplexes had been observed previously (Zhu et al. Antisense Res. Dev. 3:349-356, 1993) when an oligo(δ)ornithine moiety was covalently appended to a short oligodeoxynucleotide. We now report the analysis of duplex formation by electrophoretic gel shift analysis. In the particular example studied, an increase in Tm of 4°C was found to correspond to about a fivefold increase in binding constant. A similar enhancement by the appended cationic peptide was observed when the target strand was RNA. The use of a competitive assay format for avoidance of adsorptive loss at low concentrations (<10-7 M) of the oligonucleotide-oligo(δ)ornithine conjugate is presented.",
author = "Tianmin Zhu and Shahriar Pooyan and Ziping Wei and Leibowitz, {Michael J} and Stanley Stein",
year = "1996",
month = "3",
language = "English (US)",
volume = "6",
pages = "69--74",
journal = "Nucleic Acid Therapeutics",
issn = "2159-3337",
publisher = "Mary Ann Liebert Inc.",
number = "1",

}

TY - JOUR

T1 - Gel shift assay

T2 - Demonstration of enhanced binding of oligo(δ)-L-ornithine-oligodeoxynucleotide conjugates to complementary DNA and RNA

AU - Zhu, Tianmin

AU - Pooyan, Shahriar

AU - Wei, Ziping

AU - Leibowitz, Michael J

AU - Stein, Stanley

PY - 1996/3

Y1 - 1996/3

N2 - An increase in melting temperature for DNA:DNA duplexes had been observed previously (Zhu et al. Antisense Res. Dev. 3:349-356, 1993) when an oligo(δ)ornithine moiety was covalently appended to a short oligodeoxynucleotide. We now report the analysis of duplex formation by electrophoretic gel shift analysis. In the particular example studied, an increase in Tm of 4°C was found to correspond to about a fivefold increase in binding constant. A similar enhancement by the appended cationic peptide was observed when the target strand was RNA. The use of a competitive assay format for avoidance of adsorptive loss at low concentrations (<10-7 M) of the oligonucleotide-oligo(δ)ornithine conjugate is presented.

AB - An increase in melting temperature for DNA:DNA duplexes had been observed previously (Zhu et al. Antisense Res. Dev. 3:349-356, 1993) when an oligo(δ)ornithine moiety was covalently appended to a short oligodeoxynucleotide. We now report the analysis of duplex formation by electrophoretic gel shift analysis. In the particular example studied, an increase in Tm of 4°C was found to correspond to about a fivefold increase in binding constant. A similar enhancement by the appended cationic peptide was observed when the target strand was RNA. The use of a competitive assay format for avoidance of adsorptive loss at low concentrations (<10-7 M) of the oligonucleotide-oligo(δ)ornithine conjugate is presented.

UR - http://www.scopus.com/inward/record.url?scp=0029975621&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029975621&partnerID=8YFLogxK

M3 - Article

C2 - 8783798

AN - SCOPUS:0029975621

VL - 6

SP - 69

EP - 74

JO - Nucleic Acid Therapeutics

JF - Nucleic Acid Therapeutics

SN - 2159-3337

IS - 1

ER -