Novel nucleic acid based on small RNA interference principle

A RNA interference and nucleic acid technology, applied in DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as high cost, heavy economic burden, and poor stability of double-stranded RNA

Active Publication Date: 2015-03-04
YUNNAN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
  • Description
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Problems solved by technology

However, the stability of exogenously synthesized double-stranded RNA is poor and the cost is high, resulting in a heavy economic burden for the use of such drugs

Method used

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  • Novel nucleic acid based on small RNA interference principle
  • Novel nucleic acid based on small RNA interference principle
  • Novel nucleic acid based on small RNA interference principle

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Embodiment Construction

[0017] Add 1 mg / ml of single-stranded nucleic acid (DNA: 5`-GATTTAGCCAAGAAGTTCAGT-3`, RNA: 5`-GAUUUAGCCAAGAAGUUCAGU-3` and alternate nucleic acid dG-A-dT-U-dT-A-dG-C-dC-A -dA-G-dA-A-dG-U-dT-C-dA-G-dT, d specifically refers to deoxygenation) heat treatment at 80°C, cooled to room temperature, added liposome 2000 produced by Invitrogen Company for equal volume After mixing, prepare a series of concentrations with sterile triple distilled water after mixing. Human lung cancer A549 cells were cultured in F12K medium containing 10% fetal bovine serum at 37°C and 5% carbon dioxide. Then the cells were digested with 0.25% trypsin and adjusted to 1×10 cells with culture medium. 5 Concentration, 100 μl per well was added to a 96-well plate, and then 2 μl of serial concentrations of single-stranded nucleotides were added to each well to make the final concentration 0.91-6000 ng / ml. After 48 hours of culture, 10 μl of CCK-8 test solution was added, and the growth state of cells in each...

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Abstract

The invention relates to an alternating double strand of nucleic acid based on an RNA interference principle. Bases of the sequence of the alternating double-stranded nucleic acid consist of adenine A, guanine G, thymine T, uracil U and cytosine C. The bases are formed by alternately connecting ribose phosphate and desoxyribose phosphate. The double strand can form complete pairing or most of pairing, and each strand is 18 to 30 bases in length. The best base sequence in the first strand is 5'-GAUUUAGCCAAGAAGUUCAGU-3', and the best base sequence in the second strand is 5'-UGAACUUCUUGGCUAAAUCGC-3'. The double strand can interfere with ribonucleotide reductase so as to have an anti-tumor effect, wherein the bases U and T are replaceable.

Description

technical field [0001] The invention relates to a novel nucleic acid based on the principle of RNA interference: alternate nucleic acid double strands. The bases of its sequence are composed of adenine A, guanine G, thymine T, uracil U, and cytosine C, and each base is alternately connected by phosphoribose and phosphoribose deoxyribose. Background technique [0002] Small RNA interference is a common phenomenon in biology. Through interference, the expression level of certain proteins can be reduced at the translation level, thereby affecting biological functions. Organisms can produce double-stranded small RNAs of about 21 base pairs (bp) in a variety of ways. Small RNAs recognize specific RNAs through base pairing, and then degrade and destroy the recognized RNAs through specific mechanisms, ultimately preventing or reducing the production of functional proteins. translate. [0003] If the small RNA formed in the body is imitated in an exogenous way, it can also purpose...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113
Inventor 段为钢侯肖霖
Owner YUNNAN UNIV OF TRADITIONAL CHINESE MEDICINE
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