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Antisense oligonucleotide sequence resistant to Candida albicans infection and its uses

An antisense oligonucleotide and Candida albicans technology, which can be used in antibacterial drugs, medical preparations containing active ingredients, genetic material components, etc., and can solve problems such as strong side effects

Inactive Publication Date: 2006-02-08
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although both antibiotics and Pentamidine can inhibit the self-cleavage of type I intron ribozymes, their non-specificity of action sites in the body will lead to strong side effects in the treatment of fungal infections

Method used

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  • Antisense oligonucleotide sequence resistant to Candida albicans infection and its uses
  • Antisense oligonucleotide sequence resistant to Candida albicans infection and its uses
  • Antisense oligonucleotide sequence resistant to Candida albicans infection and its uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Screening of antisense oligonucleotides for inhibition of self-cleavage of Candida albicans type I ribozyme during in vitro transcription.

[0047] The applicant has designed 21 oligonucleotides (1-26R, 18-40R, 30-50R, 50-68R, 69-92R, 93- 109R, 107-126R, 120-138R, 137-159R, 160-179R, 180-203R, 204-220R, 221-241R, 239-262R, 255-277R, 267-288R, 280-300R, 299-320R, 317-338R, 337-359R, 356-379R, synthesized by Sebasant), they cover the entire ribozyme sequence and have the same Tm value (annealing temperature). Then, the antisense oligonucleotide that can effectively inhibit the self-cleavage system among the 21 antisense oligonucleotides was screened by the self-cleavage reaction system of type I intron ribozyme in Candida albicans 26S rRNA. The specific embodiment is: in the self-cleavage system of the type I intron ribozyme in Candida albicans 26SrRNA, the intron of the self-cleavage ribozyme is 379 nucleotides, and the 5' exon is 129 nucleotides. nucleotide...

Embodiment 2

[0056] Example 2: Confirmation of important sites in the core pseudoknot region affecting ribozyme activity

[0057] It has been determined by Example 1 that the core pseudoknot region of the type I intron ribozyme in Candida albicans 26S rRNA is very sensitive to antisense oligonucleotides, and targeting this region by antisense oligonucleotides will cause ribozyme Strong inhibition of white cleavage reactions, while RNase H cleavage experiments also showed that all oligonucleotides were bound at the correct positions. The next step is to further identify the nucleotides in the core pseudoknot region that are important for the self-cleavage activity of the ribozyme. The specific embodiment is: first fix the G of the core pseudoknot region of the I-type intron ribozyme in the Candida albicans 26S rRNA 235 site, followed by G 235 → The direction of the 3' end is designed to shorten the length of the antisense oligonucleotide; similarly fix the G of the core pseudoknot region ...

Embodiment 3

[0070] Example 3: Observation of Candida albicans absorption of antisense oligonucleotides by confocal fluorescence microscope

[0071] It has been proved by experiments that antisense oligonucleotides targeting the core pseudoknot region of type I intron ribozyme in Candida albicans 26S rRNA can cause strong inhibition of ribozyme self-cleavage reaction, but in Candida albicans Whether antisense oligonucleotides in vivo can cause strong inhibition of ribozyme self-cleavage reaction is the concern of the present invention. But how to introduce these antisense oligonucleotides targeting the core pseudoknot of Candida albicans type I ribozyme into the body of Candida albicans is a key problem. Studies have shown that Candida albicans is an excellent system for transporting antisense oligonucleotides, which can enrich antisense oligonucleotides into Candida albicans so that the concentration of antisense oligonucleotides in the body reaches 10 times that in vitro . And antisens...

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Abstract

The invention discloses an antisense oligonucleotide sequence against candida albicans infection and its use, screening an antisense oligonucleotide SEQ ID No:1 and its derivative using the core pseudoknot of candida albicans type I introne ribozyme as the targets, thus repressing the growth of the candida albicans. Test result shows that the antisense oligonucleotide can strongly repress the shearing reactivity of the candida albicans, and can result in the death of the candida albicans cells.

Description

technical field [0001] The present invention belongs to the technical field of antisense oligonucleotides, and more specifically belongs to the sequence of an antisense oligonucleotide targeting Candida albicans type I ribozyme core pseudoknot, and relates to the antisense oligonucleotide Application in the treatment of Candida albicans infection. Background technique [0002] Fungal infections, especially opportunistic fungal infections, have caused serious clinical problems in North America and some developed countries in Europe in the past two decades. The prevalence of opportunistic fungal infections has followed the rapid increase in AIDS, cancer, and organ transplantation. The immune system of AIDS patients collapses, and cancer chemotherapy and organ transplantation will cause a sharp decline in the patient's immunity. Opportunistic fungi such as Candida albicans and Pneumocystis carinii cause serious and difficult-to-treat infections in ...

Claims

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Application Information

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IPC IPC(8): C12N15/11A61K48/00A61K31/7088A61P31/04C12N15/113
Inventor 张翼张礼斌
Owner WUHAN UNIV
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