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

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 journal › Article › peer-review

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 T_m 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 language	English (US)
Pages (from-to)	69-74
Number of pages	6
Journal	Antisense and Nucleic Acid Drug Development
Volume	6
Issue number	1
State	Published - Mar 1996
Externally published	Yes

ASJC Scopus subject areas

Genetics
Pharmacology

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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 journal › Article › peer-review

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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.",

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AU - Leibowitz, Michael J

AU - Stein, Stanley

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

Abstract

ASJC Scopus subject areas

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