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Application of circular DNA (Deoxyribonucleic Acid) in detecting and imaging sense RNA (Ribonucleic Acid) and gene therapy

A technology of gene therapy and DNA probe, applied in the biological field, can solve the problems of unseen detection, imaging and gene therapy, etc., and achieve the effect of high signal-to-noise ratio, low false positive signal, and high-efficiency biological detection

Pending Publication Date: 2018-05-04
FUZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been many studies on the synthesis and properties of circular DNA, it has not been applied to detection, imaging and gene therapy in vivo

Method used

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  • Application of circular DNA (Deoxyribonucleic Acid) in detecting and imaging sense RNA (Ribonucleic Acid) and gene therapy
  • Application of circular DNA (Deoxyribonucleic Acid) in detecting and imaging sense RNA (Ribonucleic Acid) and gene therapy
  • Application of circular DNA (Deoxyribonucleic Acid) in detecting and imaging sense RNA (Ribonucleic Acid) and gene therapy

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Preparation of circular DNA: It is formed by ssDNA self-ligation and circularization. After adding primer sequence as an initiator to form a local double-stranded structure, react overnight at 16°C under the catalysis of T4 DNA ligase, and then place it at 65°C for 10 minutes to make the ligase Inactivate. Then exonuclease I and III were added to react at 37°C for 1h to degrade the unreacted ssDNA and primer sequences, and then heated to 90°C for 10min to inactivate the enzyme. Finally, the product was purified with the QIAEX II Gel Recovery Kit (Qiagen) to obtain pure circular DNA, which was quantified by measuring the absorbance at 260 nm with a UV-Vis spectrophotometer.

[0048] In Example 1, Survivin mRNA was used as the target sense RNA, and the sequences of the corresponding ssDNA and sense RNA are shown in Table 1.

Embodiment 2

[0055] use c-raf mRNA is used as the target sense RNA, and the preparation of the corresponding circular DNA is the same as that in Example 1, and the sequences of the corresponding ssDNA and sense RNA are shown in Table 2.

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Abstract

The invention provides application of circular DNA (Deoxyribonucleic Acid) in detecting and imaging sense RNA (Ribonucleic Acid) and gene therapy. The circular DNA at least comprises one or more DNA probe fragments complementary to a detection target sense RNA sequence, and is formed by connecting and cyclizing 3' and 5' terminals of ssDNA (single-stranded Deoxyribonucleic Acid). The circular DNAand the detection target sense RNA form heterozygous double strands, and the heterozygous double strands are separated from a carrier, so that (1) the application in detecting and imaging through a corresponding fluorescence signal change is realized; (2) the sense RNA in the heterozygous double strands is enzymically decomposed, so that the aim of gene therapy is achieved.

Description

technical field [0001] The invention relates to the application of circular DNA in the field of detection, imaging and gene therapy, in particular to the application of circular DNA in positive RNA detection, imaging and gene therapy, and belongs to the field of biotechnology. Background technique [0002] In addition to being the genetic material of organisms, DNA has been considered as an ideal structural unit for the construction of molecular probes in recent years, and is widely used in the fields of bioanalysis and biomedicine, including nucleic acid aptamers, molecular beacons and DNAzymes. A functionalized DNA is used in biosensing, diagnosis and disease treatment. DNA has the advantages of simple synthesis, suitable for structural modification, high selectivity and affinity, but at the same time, the biggest obstacle to the application of DNA in living organisms is that the stability and bioavailability are not high enough, resulting in protein interference and enzym...

Claims

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Application Information

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IPC IPC(8): C12Q1/6876A61K31/711A61K48/00A61P35/00
CPCA61K31/711C12Q1/6876
Inventor 李婧影李娟周洁杨黄浩
Owner FUZHOU UNIV
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