CN110144350A - The siRNA and its application in inhibition scar is formed that one species specificity inhibits CTGF gene expression - Google Patents

The siRNA and its application in inhibition scar is formed that one species specificity inhibits CTGF gene expression Download PDF

Info

Publication number
CN110144350A
CN110144350A CN201910404445.9A CN201910404445A CN110144350A CN 110144350 A CN110144350 A CN 110144350A CN 201910404445 A CN201910404445 A CN 201910404445A CN 110144350 A CN110144350 A CN 110144350A
Authority
CN
China
Prior art keywords
seq
sirna
gene expression
positive
ctgf gene
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
CN201910404445.9A
Other languages
Chinese (zh)
Inventor
王万恒
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jinuotaikang Biotechnology (beijing) Co Ltd
Original Assignee
Jinuotaikang Biotechnology (beijing) Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Jinuotaikang Biotechnology (beijing) Co Ltd filed Critical Jinuotaikang Biotechnology (beijing) Co Ltd
Priority to CN201910404445.9A priority Critical patent/CN110144350A/en
Publication of CN110144350A publication Critical patent/CN110144350A/en
Withdrawn legal-status Critical Current

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • A61P17/02Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/11DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1136Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against growth factors, growth regulators, cytokines, lymphokines or hormones
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/10Type of nucleic acid
    • C12N2310/14Type of nucleic acid interfering N.A.

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Endocrinology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Plant Pathology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dermatology (AREA)
  • Biochemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The siRNA for inhibiting CTGF gene expression the invention discloses a species specificity and its application in inhibition scar is formed.The siRNA includes the positive-sense strand and antisense strand of complete complementary, and the nucleotides sequence of the siRNA positive-sense strand is classified as any one in SEQ ID NO:1,3,5,7,9,11,13,15,17;The nucleotides sequence of the siRNA antisense strand is classified as corresponding with positive-sense strand SEQ ID NO:2,4,6,8,10,12,14,16,18.SiRNA molecule of the invention being capable of the specific expression for inhibiting CTGF gene in humans and animals cell, inhibition or the formation for reducing scar.

Description

One species specificity inhibits the siRNA of CTGF gene expression and its scar is being inhibited to be formed In application
Technical field
The present invention relates to field of biotechnology, a more specifically species specificity inhibit CTGF gene expression siRNA and Its application in inhibition scar is formed.
Background technique
China sees a doctor because of wound be up to 62,000,000 people every year, disfiguring keliod patients up to 12,000,000 people, the whole nation 2014 Hospitalisation for surgery person-time is 4382.92 ten thousand person-times;Human skin wound/postwar wound, incised injury, burn, skin infection, surgery hand Necessarily lead to scar during the skin wound healings such as art, scar be normal skin tissue's mode of appearance caused by above-mentioned reason and Tissue pathologies change.Wound healing process abnormal reaction, hyperplasia form pathologic scar.And neck surface and extremities joint The pathologic scar at position not only influences appearance and even disfeatures, it is also possible to cause dysfunction, be often accompanied by itch, pain, infection Etc. complication, be to burn one of most common complication after (wound) wound, operation, bring great damage to patient's physical and mental health, be to create One of the problem that traumatology, plastic surgery and dermatology face.
Currently, effective single therapy scheme has not been established in treatment scar or prevention hand postoperative scar growth, it is existing Treatment method mainly have physical therapy, operation excision and the common drug treatments such as steroids/chemotherapeutics/interferon and Growth factor associated medication therapies.Physical therapy is to inhibit and treat scar by the modes such as such as electricity, light, magnetic, sound, pressure, is treated Effect and risk are simultaneously deposited.Operation excision uses when often influencing dysfunction for pathologic scar, and recurrence rate is higher;Steroids is swashed For the drug therapies such as element, long-time service Patients Treated with Steroid will cause corticosteroid increase disease, pigmentation, skin and burst The adverse reactions such as ulcer, and high recurrence rate;Chemotherapeutics toxicity can lead to anaemia, thrombopenia and leukopenia etc.. Different degrees of negative effect can be all brought to patient for above-mentioned four kinds for the treatment of methods and treatment is often after cicatrization, curative effect compared with Difference.And growth factor associated medication therapies are then the generations by blocking the conduction of cytokine signalling pathways to inhibit scar, It plays a role before cicatrization, to reduce the formation of scar, plays the role for the treatment of.Such as fibroblast growth factor (FGF), a- tumor necrosis factor (TNF-a), gamma interferon (IFN-γ), epidermal growth factor (EGF), β transforming growth factor (TGF-β) etc..Though thering is artificial synthesized transforming growth factor TGF-β avotermin (avotermin) to enter the III phase at present to face The bed experimental stage, but growth factor related drugs are still less in the application study of scar treatment.
Growth factor plays an important role in Process of Forming Scar, and TGF-β 1 is fibrotic disease occurrence and development process In most important virulence factor.Connective Tissue Growth Factor (connective tissue growth factor, CTGF) is one Kind promotees the growth factor of fibrosis, passes through and promotes cell Proliferation, adjusts the biological effect of the effects of Extracellular Matrix Gene Expression performance It answers, can be seldom detected in adult normal skin histology.CTGF is the direct downstream effect medium of TGF-β 1, is stimulated into fibre Dimension cell Proliferation and the extrtacellular matrix deposition based on collagen, the fibroblast of proliferation constantly secrete CTGF, eventually lead to Fibrosis promotes pathologic scar to be formed.And the biological function of CTGF is more single, specificity is high, its expression is blocked to be not easy to draw Play some problems such as wound healing delay, inflammatory reaction.Sisco M's in 2008 etc. studies have shown that in CTGF experiment in vitro 90%mRNA is suppressed, the experiment in internal cicatricial tissue, and 55%CTGF mRNA expression is suppressed, and substantially reduces CTGF albumen Expression 20%, reduces the tendon fibroblasts quantity of the about 75% α-SMA positive, while significantly inhibiting Col I, Col III, Fibronectin (Fn) and TIMP-1mRNA expression, and finally limitation inhibits cicatrization, while not influencing wound healing.
SiRNA (small interfering RNA, siRNA) is the short sequence double-stranded RNA point of 21~25nt size Son.Transfection and the homologous siRNA of target gene to target cell, are combined by complementary with the single-stranded mRNA of target gene, start target base Because of program of degrading, the mRNA of target sequence is induced to degrade, blocks expression of target gene, the defect phenotype of cells show specific gene.With Conventional antisense nucleic acid is compared, and siRNA is technically simple easy, has gene inhibition rate is high, high specificity and concentration are low etc. Advantage is widely used in gene functional research field, provides base for tumour, genetic disease and virus infection etc. Because of the new way for the treatment of.
Summary of the invention
The present invention is exactly that in order to solve the above-mentioned technical problem, the species specificity proposed inhibits CTGF gene expression SiRNA and its application in inhibition scar is formed.
The present invention is realized according to following technical scheme.
The siRNA of one species specificity inhibition CTGF gene expression, it is characterised in that: the siRNA includes complete complementary The nucleotides sequence of positive-sense strand and antisense strand, the siRNA positive-sense strand is classified as SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35、SEQ ID Any one in NO:37;The nucleotides sequence of the siRNA antisense strand is classified as SEQ ID NO corresponding with positive-sense strand: 22、SEQ ID NO:24、SEQ ID NO:26、SEQ ID NO:28、SEQ ID NO:30、SEQ ID NO:32、SEQ ID NO:34、SEQ ID NO:36、SEQ ID NO:38。
Further, at least one 3 ' single-stranded ends are also connected with 1 to 3 in the positive-sense strand and antisense strand of the siRNA A additional nucleotide, to form at least one after siRNA positive-sense strand and antisense strand complementary pairing by 1 to 3 nucleotide 3 ' the jags constituted.
Further, the 3 ' jag is continuous 2 uridylate UU.
Further, the siRNA positive-sense strand and antisense strand all contain 3 ' jags.
One species specificity inhibits the siRNA of CTGF gene expression, and the siRNA includes the positive-sense strand and antisense of complete complementary The nucleotides sequence of chain, the siRNA positive-sense strand is classified as SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID It is NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, any in SEQ ID NO:17 One;The nucleotides sequence of the siRNA antisense strand be classified as SEQ ID NO:2 corresponding with positive-sense strand, SEQ ID NO:4, SEQ ID NO:6、SEQ ID NO:8、SEQ ID NO:10、SEQ ID NO:12、SEQ ID NO:14、SEQ ID NO:16、 SEQ ID NO:18。
It is a kind of it is above-mentioned specificity inhibit CTGF gene expression siRNA inhibit scar formed in purposes.
Further, the siRNA inhibits the expression of CTGF gene in humans and animals cell by specificity, to inhibit Or reduce the formation of scar.
Present invention obtains following beneficial effects.
The invention discloses the nucleotide sequences (siRNA) of nine groups of RNA interferings for CTGF gene expression, being capable of target To inhibition CTGF gene expression and then it is used to treat disease relevant to CTGF gene unconventionality expression.In addition, of the present invention SiRNA sequence has high homology in different plant species, due to this high homology, on the one hand, between different plant species Test can use mutually homotactic siRNA, reduce the mistake for constituting and synthesizing different sequence siRNA according to the gene of different plant species Journey can greatly shorten experiment process when test different animals model, various preclinical tests are rapidly completed;Another party Face, also can avoid using brought by different sequence siRNA to corresponding animal CTGF mRNA inhibit efficiency, stability not really It is qualitative, so as to accelerate Subsequent pharmacological to research and develop process.
Detailed description of the invention
Fig. 1 is the Alexa that inverted fluorescence microscope of the present invention transfects triumphant peptideRed Fluorescent Transfected condition figure of the Oligo in three kinds of cell lines;
Fig. 2 is influence diagram of the transfection reagent of the present invention to three kinds of Apoptosis;
Fig. 3 is present invention difference CTGF siRNA (CT1, CT2, CT3, CT9) and its concentration (1,10,100nM) to CTGF The influence diagram of expression.
Specific embodiment
The invention will be further described with reference to the accompanying drawings and embodiments.
Embodiment 1siRNA sequence information
With CTGF mRNA, (Genebank number of registration: the template that (NM_001901) designs for siRNA obtains three to the present invention The conservative siRNA of species, sequence information are shown in Table 1.Meanwhile positive-sense strand nucleotide sequence is set as shown in sequence 19 and antisense Chain the nucleotide sequence siRNA as shown in sequence 20, number NC.NC is the nothing with CTGF mRNA without corresponding target action site Sequence is closed, as negative control.
The siRNA information of 1: three, table targeting CTGF
The positive-sense strand of listed siRNA and antisense strand are synthesized by Shanghai Ji Ma Bioisystech Co., Ltd in table 1.Wherein, Sequence SEQ ID NO:1-SEQ ID NO:18 is corresponding in turn to for the 3 ' end sequence SEQ ID NO:21-SEQ ID NO:38 to be drawn Enter 2 uridylates and is formed.Sequence SEQ ID NO:1-SEQ ID NO:18 is equimolar with annealing salt solution Positive-sense strand and antisense strand mixture, for subsequent conventional annealing to form siRNA double-strand, it is 3 ' prominent to be respectively provided with UU for the both ends of double-strand Outlet.
Embodiment 2: transfection reagent verifying
Triumphant peptide gene transfection reagent (Kai Er Bioisystech Co., Ltd, Shanxi China Telecom) is transfection reagent used in the present invention, this Invention first verifies the transfection efficiency and cytotoxicity of triumphant peptide transfection siRNA, excludes influence of the transfection reagent to experiment.
(1) cell transfecting efficiency
Human lung carcinoma cell (maxicell lung cancer) (H460 is purchased from ATCC, BNCC102112), gland cancer alveolar substrate epithelium is thin Born of the same parents' (A549 cell) (be purchased from ATCC, BNCC100258) and human bronchial epithelial like cell (HBE, purchased from ATCC, BNCC338600) cell inoculation is in 24 orifice plates, inoculum density 105Cells/well, with the cell containing F12+10% fetal calf serum Culture medium is placed in 37 DEG C of 5%CO2Culture.Cell transfecting illustrates the BLOCK- for transfecting 50nM respectively according to triumphant peptide gene transfection reagent iTTMAlexaRed Fluorescent Oligo (be purchased from invitrogen) arrives H460, A549 and HBE cell, and 37 DEG C Culture.Pass through inverted fluorescence microscope and flow cytomery cell transfecting efficiency afterwards for 24 hours, the results showed that in three kinds of cells The transfection efficiency of triumphant peptide transfection siRNA is all larger than 90% (Fig. 1) in system.
(2) cytotoxicity
By H460, A549 and HBE cell inoculation in 24 orifice plates, inoculum density 105Cells/well, with containing F12+10% The cell culture medium of fetal calf serum is placed in 37 DEG C of 5%CO2Overnight incubation.It is greater than 90% triumphant peptide reagent amount with transfection efficiency respectively Handle A549, H460 and HBE cell, 37 DEG C of 5%CO2Cultivate 72h.Respectively for 24 hours, 48h and 72h pass through Annexin V- EGFP/PI Apoptosis Detection Kit (be purchased from Beijing Quanshijin Biotechnology Co., Ltd) and fluorescence microscope or Flow cytomery cell survival rate.Negative control is the cell culture medium of F12+10% fetal calf serum, F12 cell culture fluid (being purchased from GIBCO), fetal calf serum (are purchased from Biological Industries), and reference reagent is Lipofectaine RNAiMAX (is purchased from Invitrogen).The result shows that cell and untransfected of the cell transfecting efficiency in 90-99%, after transfection Cell compare, Apoptosis there are no significant difference shows triumphant peptide transfection reagent almost no cytotoxicity (Fig. 2).
Embodiment 3: the real-time PCR of fluorescent quantitation (qRT-PCR) detects siRNA in vitro to CTGF mRNA expression Inhibitory effect.
By A549 cell inoculation in 24 orifice plates, inoculum density 105Cells/well, the cell training of F12+10% fetal calf serum Feeding base is placed in 37 DEG C of 5%CO2Overnight incubation.
Illustrate to transfect 4 kinds of CTGF siRNA (CT1:SEQ ID NO:1-SEQ ID respectively according to triumphant peptide gene transfection reagent NO:2;CT2:SEQ ID NO:3-SEQ ID NO:4;CT3:SEQ ID NO:5-SEQ ID NO:6;CT9:SEQ ID NO: 17-SEQ ID NO:18) He Yiwu targeting negative control siRNA (being denoted as NC) arrive A549 cell, siRNA concentration be 1,10 And 100nM, then in 37 DEG C of 5%CO2Cultivate 48h.
After the cell 48h of culture transfection, the total serum IgE in cell is extracted with Rneasy Mini Kit (Qiagen company). Each sample takes 2 μ g total serum IgEs, according to PrimeScriptTM1st Strand cDNA Synthesis Kit (Takara company) Application method reverse transcription obtain cDNA, using TaqMan Assay detection kit (Applied Biosystems company) into Row fluorescent quantitation real-time PCR reactions.PCR condition is as follows: 95 DEG C of initial denaturation 10min, and into circulation: 95 DEG C of denaturation 30s, 60 DEG C are moved back Fiery 30s, 72 DEG C of extension 30s, after 40 recycle, 72 DEG C of extension 10min.User β 2microglobulin (B2M) is as quantitative The reference gene (Applied Biosystems company, Catalog number:4310886E) of PCR reaction, the base of CTGF mesh Because the Ct value of expression is standardized with the Ct value of B2M, the calculating of the multiple variation of sample amplification uses 2–△△tMethod.PCR used It expands CTGF primer and B2M primer sequence is as shown in table 3.
Table 3: quantitative PCR detection primer sequence
Title Upstream (5 ' -3 ') Downstream (5 ' -3 ')
People-CTGF CACCCGGGTTACCAATGACA TCCGGGACAGTTGTAATGGC
The results show that the cell through handling 48h, CT9 and CT3 have significant inhibiting effect to the expression of CTGF mRNA, with The expression of increase (1nM, 10nM, 100nM) the CTGF mRNA of its siRNA dosage gradually decrease.Such as 1nM CT9siRNA couple CTGF expression inhibiting rate is that the processing of 36%, 100nM can be promoted to 88%, and the processing of negative control siRNA is to the table of CTGF Up to not influencing, show that CTGF siRNA transfects the expression (Fig. 3) for successfully and effectively having suppressed CTGF.
Sequence table
<110>Jino Tai Kang biotechnology (Beijing) Co., Ltd
The siRNA and its application in inhibition scar is formed that<120>one species specificity inhibit CTGF gene expression
<160> 38
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
ccggguuacc aaugacaacu u 21
<210> 2
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 2
guugucauug guaacccggu u 21
<210> 3
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 3
agaccugugg gaugggcauu u 21
<210> 4
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 4
augcccaucc cacaggucuu u 21
<210> 5
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 5
ccgacuggaa gacacguuuu u 21
<210> 6
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 6
aaacgugucu uccagucggu u 21
<210> 7
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 7
gcaccagcau gaagacauau u 21
<210> 8
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 8
uaugucuuca ugcuggugcu u 21
<210> 9
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 9
gaagacacgu uuggcccagu u 21
<210> 10
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 10
cugggccaaa cgugucuucu u 21
<210> 11
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 11
gcuaaauucu guggaguauu u 21
<210> 12
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 12
auacuccaca gaauuuagcu u 21
<210> 13
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 13
gcgaggucau gaagaagaau u 21
<210> 14
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 14
uucuucuuca ugaccucgcu u 21
<210> 15
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 15
aagcugaccu ggaagagaau u 21
<210> 16
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 16
uucucuucca ggucagcuuu u 21
<210> 17
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 17
ccuaucaagu uugagcuuuu u 21
<210> 18
<211> 21
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 18
aaagcucaaa cuugauaggu u 21
<210> 19
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 19
uucuccgaac gucgcacgu 19
<210> 20
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 20
acgugacacg uucggagaa 19
<210> 21
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 21
ccggguuacc aaugacaac 19
<210> 22
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 22
guugucauug guaacccgg 19
<210> 23
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 23
agaccugugg gaugggcau 19
<210> 24
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 24
augcccaucc cacaggucu 19
<210> 25
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 25
ccgacuggaa gacacguuu 19
<210> 26
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 26
aaacgugucu uccagucgg 19
<210> 27
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 27
gcaccagcau gaagacaua 19
<210> 28
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 28
uaugucuuca ugcuggugc 19
<210> 29
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 29
gaagacacgu uuggcccag 19
<210> 30
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 30
cugggccaaa cgugucuuc 19
<210> 31
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 31
gcuaaauucu guggaguau 19
<210> 32
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 32
auacuccaca gaauuuagc 19
<210> 33
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 33
gcgaggucau gaagaagaa 19
<210> 34
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 34
uucuucuuca ugaccucgc 19
<210> 35
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 35
aagcugaccu ggaagagaa 19
<210> 36
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 36
uucucuucca ggucagcuu 19
<210> 37
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 37
ccuaucaagu uugagcuuu 19
<210> 38
<211> 19
<212> RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 38
aaagcucaaa cuugauagg 19

Claims (7)

1. a species specificity inhibition CTGF gene expression siRNA, it is characterised in that: the siRNA include complete complementary just The nucleotides sequence of adopted chain and antisense strand, the siRNA positive-sense strand is classified as SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35、SEQ ID Any one in NO:37;The nucleotides sequence of the siRNA antisense strand is classified as SEQ ID NO corresponding with positive-sense strand: 22、SEQ ID NO:24、SEQ ID NO:26、SEQ ID NO:28、SEQ ID NO:30、SEQ ID NO:32、SEQ ID NO:34、SEQ ID NO:36、SEQ ID NO:38。
2. the siRNA that a species specificity according to claim 1 inhibits CTGF gene expression, it is characterised in that: described At least one 3 ' single-stranded ends are also connected with 1 to 3 additional nucleotide in the positive-sense strand and antisense strand of siRNA, thus At least one 3 ' jag being made of 1 to 3 nucleotide is formed after siRNA positive-sense strand and antisense strand complementary pairing.
3. the siRNA that a species specificity according to claim 2 inhibits CTGF gene expression, it is characterised in that: described 3 ' Jag is continuous 2 uridylate UU.
4. the siRNA that a species specificity according to claim 2 inhibits CTGF gene expression, it is characterised in that: described SiRNA positive-sense strand and antisense strand all contain 3 ' jags.
5. a species specificity inhibition CTGF gene expression siRNA, it is characterised in that: the siRNA include complete complementary just Adopted chain and antisense strand, the nucleotides sequence of the siRNA positive-sense strand are classified as SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO: 5、SEQ ID NO:7、SEQ ID NO:9、SEQ ID NO:11、SEQ ID NO:13、SEQ ID NO:15、SEQ ID NO:17 In any one;The nucleotides sequence of the siRNA antisense strand is classified as SEQ ID NO:2 corresponding with positive-sense strand, SEQ ID NO:4、SEQ ID NO:6、SEQ ID NO:8、SEQ ID NO:10、SEQ ID NO:12、SEQ ID NO:14、SEQ ID NO:16、SEQ ID NO:18。
6. specificity described in a kind of claim 1-5 inhibits purposes of the siRNA of CTGF gene expression in inhibition scar is formed.
7. a species specificity according to claim 6 inhibits the siRNA of CTGF gene expression in inhibition scar is formed Purposes, which is characterized in that the siRNA inhibits the expression of CTGF gene in humans and animals cell by specificity, to inhibit Or reduce the formation of scar.
CN201910404445.9A 2019-05-15 2019-05-15 The siRNA and its application in inhibition scar is formed that one species specificity inhibits CTGF gene expression Withdrawn CN110144350A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910404445.9A CN110144350A (en) 2019-05-15 2019-05-15 The siRNA and its application in inhibition scar is formed that one species specificity inhibits CTGF gene expression

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910404445.9A CN110144350A (en) 2019-05-15 2019-05-15 The siRNA and its application in inhibition scar is formed that one species specificity inhibits CTGF gene expression

Publications (1)

Publication Number Publication Date
CN110144350A true CN110144350A (en) 2019-08-20

Family

ID=67595543

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910404445.9A Withdrawn CN110144350A (en) 2019-05-15 2019-05-15 The siRNA and its application in inhibition scar is formed that one species specificity inhibits CTGF gene expression

Country Status (1)

Country Link
CN (1) CN110144350A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114807127A (en) * 2021-01-19 2022-07-29 陈璞 Small interfering RNA for connective tissue growth factor and application thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101160138A (en) * 2004-12-23 2008-04-09 爱尔康公司 Rnai inhibition of ctgf for treatment of ocular disorders
WO2013148159A1 (en) * 2012-03-30 2013-10-03 Fibrogen Inc Therapeutic methods for peritoneal carcinomatosis
CN105462975A (en) * 2015-12-18 2016-04-06 中国科学院北京基因组研究所 Small interfering RNA for specific inhibition of CTGF gene expression and application thereof
CN106047879A (en) * 2016-08-18 2016-10-26 广州市锐博生物科技有限公司 Oligonucleotide molecule used for inhibiting expression of mRNA of target gene and composition set thereof
CN108251420A (en) * 2016-12-28 2018-07-06 苏州瑞博生物技术有限公司 Inhibit siRNA, the composition comprising it and its application of CTGF gene expressions in humans and animals

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101160138A (en) * 2004-12-23 2008-04-09 爱尔康公司 Rnai inhibition of ctgf for treatment of ocular disorders
WO2013148159A1 (en) * 2012-03-30 2013-10-03 Fibrogen Inc Therapeutic methods for peritoneal carcinomatosis
CN105462975A (en) * 2015-12-18 2016-04-06 中国科学院北京基因组研究所 Small interfering RNA for specific inhibition of CTGF gene expression and application thereof
CN106047879A (en) * 2016-08-18 2016-10-26 广州市锐博生物科技有限公司 Oligonucleotide molecule used for inhibiting expression of mRNA of target gene and composition set thereof
CN108251420A (en) * 2016-12-28 2018-07-06 苏州瑞博生物技术有限公司 Inhibit siRNA, the composition comprising it and its application of CTGF gene expressions in humans and animals

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
罗媛元等: "SiRNA在肝纤维化基因治疗中的应用进展", 《山东医药》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114807127A (en) * 2021-01-19 2022-07-29 陈璞 Small interfering RNA for connective tissue growth factor and application thereof

Similar Documents

Publication Publication Date Title
Liang et al. Systematic analyses reveal long non-coding RNA (PTAF)-mediated promotion of EMT and invasion-metastasis in serous ovarian cancer
Nakasa et al. Expression of microRNA‐146 in rheumatoid arthritis synovial tissue
Chen et al. MiR-351 transiently increases during muscle regeneration and promotes progenitor cell proliferation and survival upon differentiation
JP2009522303A (en) SiRNA compositions that promote skin wound healing without scarring and methods of wound treatment
CN111035648B (en) Application of long-chain non-coding RNAGAS5 in preparation of medicine for promoting nerve regeneration and repairing nerve injury
Sullivan et al. Spatiotemporal microRNA profile in peripheral nerve regeneration: miR-138 targets vimentin and inhibits Schwann cell migration and proliferation
Robinson et al. MicroRNA signature in wound healing following excimer laser ablation: role of miR-133b on TGFβ1, CTGF, SMA, and COL1A1 expression levels in rabbit corneal fibroblasts
Guo et al. MicroRNA expression signature and the therapeutic effect of the microRNA‑21 antagomir in hypertrophic scarring
He et al. MicroRNA-181b expression in prostate cancer tissues and its influence on the biological behavior of the prostate cancer cell line PC-3
CN111317820B (en) Use of splicing factor PRPF31 inhibitor for preparing medicine
CN110903348B (en) Small peptide for promoting wound healing and application thereof
CN107142310B (en) Screening method of specific shRNA for inhibiting lung cancer cells by targeting Ang-2 gene
CN105861551B (en) The carrier and its construction method of Combined expression microRNAs inhibition Cells Proliferation of Human Breast Cancer and application
CN110144350A (en) The siRNA and its application in inhibition scar is formed that one species specificity inhibits CTGF gene expression
US11542502B2 (en) Application of microRNA-210 inhibitor in preparation of drugs for treating inflammatory skin diseases
CN113388674B (en) Application of miRNA as drug target for detecting or treating benign prostatic hyperplasia
CN110279706B (en) Application of hnRNPC gene C1 and/or C2 subtype as drug target in screening anti-cancer drugs
CN114099536A (en) Application of agent for down-regulating Czib gene expression in preparation of drugs for treating or improving pulmonary fibrosis
CN112725436A (en) Application of human circMKLN1 gene and related product
CN111575381A (en) Novel use of biomarkers
McFarland et al. Expression of genes encoding antimicrobial proteins and members of the toll‐like receptor/nuclear factor‐κB pathways in engineered human skin
CN107723367A (en) Application of miRNA-885-3p and product using same
CN111118154B (en) Application of LINC01272 in preparation of tumor detection reagent and/or treatment drug
US10443059B2 (en) Pharmaceutical composition and method for reducing scar formation
US20230279393A1 (en) Treatment of Airway Conditions by Modulation of MiR200 Family MicroRNAs

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
TA01 Transfer of patent application right
TA01 Transfer of patent application right

Effective date of registration: 20200728

Address after: Room 101, 1 / F, building 3, yard 18, Hongda South Road, Daxing District, Beijing

Applicant after: Beijing Laixin Biotechnology Co., Ltd

Address before: 100102 Beijing Chaoyang District Chaowai Avenue Yijing Garden Beili Building 15B

Applicant before: Jinuotaikang Biotechnology (Beijing) Co.,Ltd.

WW01 Invention patent application withdrawn after publication
WW01 Invention patent application withdrawn after publication

Application publication date: 20190820