CN110305865A - A kind of siRNA and its inhibition cell Proliferation for interfering ZNF24 gene expression and the application in migration - Google Patents
A kind of siRNA and its inhibition cell Proliferation for interfering ZNF24 gene expression and the application in migration Download PDFInfo
- Publication number
- CN110305865A CN110305865A CN201910602849.9A CN201910602849A CN110305865A CN 110305865 A CN110305865 A CN 110305865A CN 201910602849 A CN201910602849 A CN 201910602849A CN 110305865 A CN110305865 A CN 110305865A
- Authority
- CN
- China
- Prior art keywords
- znf24
- sirna
- gene expression
- cell
- interfering
- 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.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
- C12N2310/141—MicroRNAs, miRNAs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2320/00—Applications; Uses
- C12N2320/30—Special therapeutic applications
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- General Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention provides a kind of siRNA for interfering ZNF24 gene expression and its application in inhibiting cell Proliferation and migrating.Three couples of siRNA of compounding design of the present invention, are ZNF24-siRNA-292, ZNF24-siRNA-485 and ZNF24-siRNA-575 respectively, these siRNA can effectively, specifically interfere the expression of ZNF24, so as to effectively inhibit the proliferation and migration of cell.Invention increases the quantity of the siRNA sequence for interference ZNF24 gene expression, and inhibit the proliferation and migration of cell by RNA interference specificity inhibition ZNF24 gene expression in cellular level, so as to inhibit, mitigate or prevent the disease that may cause by ZNF24 gene, and lay a good foundation for mechanism of action of the research ZNF24 gene in cell Proliferation and migration with following application study.
Description
Technical field
The invention belongs to molecular genetics fields, and in particular to a kind of siRNA for interfering ZNF24 gene expression and its
Inhibit the application in cell Proliferation and migration.
Background technique
RNA interference (RNA interference, RNAi) is silencing after the homologous target gene transcription by double chain RNA mediate
Process, it can have mutually homotactic double-stranded RNA by artificial introducing and endogenous target gene, induce endogenous target gene mRNA
Degradation, to achieve the purpose that reduce gene expression.RNAi phenomenon, which is also that one kind is conservative in evolution, resists transgenosis or external
The defense mechanism that virus is invaded.Although the search time of RNA perturbation technique is shorter, it is as a kind of emerging gene disruption
Technology, has that specificity is high, jamming effectiveness is high and simple operation and other advantages, and is widely used in virus infection, cancer, dominant
In the medical researches such as hereditary disease, drug target gene can be screened with high throughput, promote gene therapy, new drug development etc., and it is small dry
It disturbs RNA (small interfering RNA, siRNA) and is also referred to as " disease medicine ".In recent years, scientist utilizes RNAi skill
Art selects the target gene of specificity to synthesize effective siRNA, and selects Stable transfection mode that siRNA transduces to specific
Into the cell, act on it efficiently in host cell, the expression of silencing related gene, this kind of means can be from genomic level point
The function of analysis gene in the mammalian body can also more quickly identify many genes relevant to disease, make us can be with
Preferably study organism in gene regulation network system, therefore RNAi technology have become research gene function it is most effective
Means, also become the most attractive method of target gene therapy.And siRNA is in mammalian cell, organ and living body
Lesion in healing function, imply in the near future, RNAi can also be used successfully to the treatment of human diseases, so related
The research and application of siRNA has extremely important theoretical and practical significance, far-reaching by generating to the development of Medical Biology
It influences.
Proliferation, migration and the invasion of cell are the molecular biology change procedures of multifactor participation, multi-step completion.It is certain
Cell with very strong transfer ability can enable it to overcome the adhesive attraction between cell and iuntercellular and cell and matrix
And invade surrounding tissue；The strong cell of invasion generally also has many synapse cells, it can make intercellular adhesion weaken and
Communication connection forfeiture etc..There is very important meaning to disease, cell application etc. accordingly, with respect to the research of cell Proliferation and migration
Justice.
Human zinc finger protein gene (ZNF24) is made of 4 exons, and coding includes the class of 368 amino acid residues
Kruppel albumen.The transcript of ZNF24 largely exists in heart, liver, lung and spleen, exists in urogenital tissue, lymph
In tissue, small intestine and colon, low-level exists in the brain.And also there is research sub- thin by Green fluorescent protein fusion vector
Born of the same parents' positioning experiment discovery ZNF24 is located on nucleus, the TCAT tetranucleotide being widely present in its energy specificity and genome
Repetitive sequence combines, and ZNF24 is the central nervous system myelin formation of oligodendroglia and maintains neural progenitor cell point
It splits necessary to state.Protein science experiment also indicates that ZNF24 is the DNA replication dna factor, has important physiological function, Ke Yican
With the regulation of kinases transcriptional activity, blood vessel hyperplasia, brain growth and DNA damage response etc..But to currently, related interference ZNF24 gene
The research of expression is fewer, and targets the application of the siRNA sequence cells certain for exploitation for interference ZNF24 gene expression
Research has important meaning.
Summary of the invention
The siRNA that the object of the present invention is to provide a kind of for interfering ZNF24 gene expression and its inhibit cell Proliferation and
Application in migration.The present invention synthesizes three couples of siRNA of targeted inhibition ZNF24 gene expression by design, and successfully constructs
Their expression vectors intracellular in HEK-293, while the table of ZNF24 can effectively, be specifically interfered with these siRNA
It reaches, to effectively inhibit the proliferation and migration of cell.
For achieving the above object, the present invention is achieved by the following scheme:
It is provided by the invention a kind of for interfering the ZNF24-siRNA-292 of ZNF24 gene expression, which is characterized in that institute
The nucleotide sequence of the ZNF24-siRNA-292 for interfering ZNF24 gene expression stated are as follows:
Positive-sense strand: 5 '-GCGAAGAGGGATCAAGTATTT-3 ',
Antisense strand: 5 '-ATACTTGATCCCTCTTCGCTT-3 '.
Provided by the invention a kind of for interfering the ZNF24-siRNA-485 of ZNF24 gene expression, described is used to interfere
The nucleotide sequence of the ZNF24-siRNA-485 of ZNF24 gene expression are as follows:
Positive-sense strand: 5 '-GCAGTTTGTTGCCATCCTATT-3 ',
Antisense strand: 5 '-TAGGATGGCAACAAACTGCTT-3 '.
Provided by the invention a kind of for interfering the ZNF24-siRNA-575 of ZNF24 gene expression, described is used to interfere
The nucleotide sequence of the ZNF24-siRNA-575 of ZNF24 gene expression are as follows:
Positive-sense strand: 5 '-GGATTTGGAGAGTGAACTTTT-3 ',
Antisense strand: 5 '-AAGTTCACTCTCCAAATCCTT-3 '.
The present invention also provides described for interfering the siRNA of ZNF24 gene expression being used to prepare inhibition cell increasing
Grow and the preparation that migrates in application.
Further, the application includes following operating procedure:
(1) described ZNF24-siRNA-292, ZNF24-siRNA-485 or ZNF24-siRNA-575 are transfected into after cell
Cracking obtains total serum IgE；
(2) the first chain cDNA is synthesized by template reverse transcription of the total serum IgE in step (1)；
(3) RT-PCR is carried out as template using the cDNA in step (2) and detects the inhibition that ZNF24-siRNA expresses ZNF24
Efficiency；
(4) the highest ZNF24-siRNA of inhibition efficiency in step (3) is transfected into cell, preparation inhibits cell Proliferation
With the preparation of migration.
Further, the concentration of ZNF24-siRNA is 20pmol in the step (1).
Further, the dosage of total serum IgE is 10ng-2 μ g in the step (2)
Further, RT-PCR reaction system is 20 μ l in the step (3), comprising: cDNA template 20-100ng, it is positive
10 μ l, RNase-free ddH of 1 μ l of primer, 1 μ l, SuperReal PreMix Plus of reverse primer2O is supplied to 20 μ l.
Further, RT-PCR response procedures in the step (3) are as follows: 95 DEG C of initial denaturation 15min；95 DEG C of denaturation 10s, 60
DEG C annealing 30s, 72 DEG C of extensions 30s, totally 40 recycle.
Compared with prior art, the present invention having the following advantages and beneficial effects:
(1) by three couples of siRNA of design synthesis targeting ZNF24 expression, they can effectively, specifically be interfered the present invention
The expression of ZNF24, and increase the quantity of the siRNA sequence for interference ZNF24 gene expression；
(2) present invention inhibits the proliferation of cell in cellular level by RNA interference specificity interference ZNF24 gene expression
And migration, it so as to inhibit, mitigate or prevent the disease that may cause by ZNF24 gene, and is research ZNF24 gene thin
The application study of born of the same parents' proliferation and mechanism of action and future in migration is laid a good foundation.
Detailed description of the invention
Fig. 1 is the comparison of ZNF24 expression conditions in difference ZNF24-siRNA transfection group cell of the invention.
Fig. 2 is siRNA interferes ZNF24 gene expression in difference ZNF24-siRNA transfection group cell of the invention 24 hours
Effect compares.
Fig. 3 is siRNA interferes ZNF24 gene expression in difference ZNF24-siRNA transfection group cell of the invention 48 hours
Effect compares.
Fig. 4 is the comparison of ability of cell proliferation situation in difference ZNF24-siRNA transfection group of the invention.
Fig. 5 is the comparison of cell migration capabilities might in difference ZNF24-siRNA transfection group of the invention.
Specific embodiment
Technical solution of the present invention is further described in detail below in conjunction with specific embodiment.
Embodiment 1: the design synthesis of RNA interfering (siRNA) target sequence
ZNF24 gene order is obtained from GenBank, it is conservative for ZNF24 gene order according to siRNA design principle
Region screening design simultaneously synthesize 3 interference target sequences, be respectively designated as ZNF24-siRNA-292, ZNF24-siRNA-485,
ZNF24-siRNA-575, while designing not is negative control (siFAM), sequence such as table 1 for the meaningless sequence of any gene
It is shown.
The design of 1 siRNA sequence of table and synthesis
Embodiment 2: cell culture and transfection
DMEM cell culture medium of the HEK-293 cell containing FBS and antibiotic is containing 5%CO under the conditions of 37 DEG C2It is permanent
It cultivates in warm incubator, when vitro growth rates reach 70-90%, is passed on 0.25% trypsin digestion.It transfects previous
It, by 4-5 × 104By cell inoculation on 24 orifice plates, 0.5ml DMEM culture medium is added in every hole in a/hole.When transfection, in 50 μ l
Plasma-free DMEM medium be added 20pmol ZNF24-siRNA, soft mixing；Simultaneously with 50 μ l plasma-free DMEM mediums
2000 reagent of Lipofectamin for diluting 1 μ l, is placed at room temperature for 5 minutes after mixing gently.Then by the siRNA diluted and
It is softly mixed after transfection reagent mixing, 20 minutes is placed at room temperature for, to form 2000 compound of siRNA/Lipofectamin.
The compound of 100 μ l is added in 24 orifice plate containing cell, softly rocks tissue culture plate back and forth, cell is containing 5%CO2It is permanent
It is incubated for 24 hours after -48h for 37 DEG C in warm incubator, collects other detections after cell is transfected.If cell strain is more sensitive,
Then after being incubated for 4h-6h, removes compound and replace culture medium.
The inhibition Efficiency testing of embodiment 3:siRNA
(1) Total RNAs extraction
TRNzol reagent lytic cell in the hole 1ml/ is added directly in culture plate, several times with sampler piping and druming, sample will be homogenized
Product are in 15-30 DEG C of placement 5min, so that nucleic acid-protein compound is kept completely separate.0.2ml chlorine is added in every TRNzol using 1ml
It is imitative, pipe lid is covered, 15s is acutely vibrated, after being placed at room temperature for 3min, is placed in 4 DEG C of centrifuges and is centrifuged 10- using 12,000rpm
15min, sample can be divided into three layers at this time: the organic phase of yellow, the colourless water phase of middle layer and upper layer, RNA mainly in water phase,
Water phase (about 600 μ l) is transferred in new centrifuge tube.Isometric isopropanol is added in obtained aqueous phase solution, mixes, room
4 DEG C of 12,000rpm are centrifuged 10min after temperature places 20-30min, remove supernatant.75% ethanol washing of 1ml precipitating is added, and in 4 DEG C
5,000rpm centrifugation 3min, remove supernatant.After precipitating room temperature is placed to dry, 30-100 μ l RNase-free ddH is added2O, instead
Multiple piping and druming mixes, sufficiently dissolution RNA.
(2) reverse transcription synthesizes the first chain cDNA
Template ribonucleic acid is thawed on ice；5×gDNA Buffer,FQ-RT Primer Mix,10×Fast RT
Buffer and RNase-Free ddH2O is immediately placed on ice after thaw at RT, defrosting respectively.By every kind of solution whirlpool before use
Rotation oscillation mixes, and brief centrifugation remains in the liquid of tube wall to collect.Following operating procedure is completed on ice.According to table 2
Genomic DNA removes system and prepares mixed liquor, thoroughly mixes.Brief centrifugation is placed in 42 DEG C of incubation 3min, then puts on ice
It sets.
2 gDNA of table removes reaction system
Mixed liquor is prepared according to the reverse transcription reaction system of table 3.Mixed liquor in reverse transcription reaction is added to gDNA removal
In the reaction solution of step, mix well.After 42 DEG C of incubation 15min, cDNA is obtained after 95 DEG C of incubation 3min, and be put in
Cryo-conservation.
3 reverse transcription reaction system of table
(3) fluorescent quantitative PCR experiment
Reaction solution is configured on ice according to the RT-PCR reaction system of table 4, all reagents is mixed at room temperature and thorough
It mixes.
4 RT-PCR reaction system of table
Reaction system is placed in fluorescence quantitative PCR instrument, is configured according to the RT-PCR response procedures of table 5, is started anti-
It answers.
5 RT-PCR response procedures of table
RT-PCR detects effectiveness results such as Fig. 1-3 of siRNA interference ZNF24 gene expression.As the result is shown with control group phase
Comparing, the ZNF24 gene mRNA expression amount in three ZNF24-siRNA transfection groups for 24 hours with 48h is below control group, wherein
ZNF24-siRNA-575 group most, the followed by ZNF24-siRNA-292 that ZNF24 gene expression dose is reduced in 48h, by
This illustrates that jamming effectiveness highest of the ZNF24-siRNA-575 in 48h to ZNF24 gene, effect are best.
Embodiment 4: cell proliferation experiment
The cell for taking transfection ZNF24-siRNA-575 to finish, in the every 100 μ l cell suspension of Kong Zhongjia of 96 orifice plates, wherein setting 6
A multiple holes are containing 5%CO2Constant incubator in 37 DEG C of overnight incubations；After culture 1-7 days, every hole adds 10 μ l CCK8 dye liquors,
Containing 5%CO2Constant incubator in 37 DEG C of culture 2h；The absorbance value in every hole under 450nm wavelength is detected with microplate reader
(OD450)；It is mapped with absorbance value to time (number of days), draws cell Proliferation curve.
CCK-8 detects the result of cell Proliferation as shown in figure 4, compared with the control group, cell after ZNF24 expression silencing
Proliferative capacity gradually weakens over time, illustrates that siRNA interference ZNF24 gene expression is able to suppress cell Proliferation energy
Power.
Embodiment 5: cell migration assay
The effective disturbance target point ZNF24-siRNA-575 cell of selection siRNA and cellular control unit are cultivated, and logarithmic phase is taken
Transfection siRNA cell, conventional digestion, centrifugation abandon supernatant after, be added plasma-free DMEM medium be resuspended cell, prepare slender
Density is adjusted to 5 × 10 by born of the same parents' suspension5A cell/ml.It is outstanding that 200 μ l cells are added into the upper layer of each cell Transwell
DMEM culture medium of the 700 μ l containing 10%FBS is added along lower layer's inner wall in liquid, avoids generating bubble, is containing 5%CO2Constant temperature training
Support 37 DEG C of incubation 16h in case.The cell Transwell is taken out, the cell of cell filter membrane upper surface is carefully wiped with cotton swab, then will be small
Room, which is put into methanol, fixes 15min, cleans 3 times with PBS to remove remaining methanol, is put into 0.2% crystal violet dye liquor and contaminates
Color 20min, then 3 times are cleaned with PBS to remove extra dyeing liquor, it is then placed within microscopically observation and takes pictures.
The result of Transwell testing inspection cell migration is as shown in figure 5, compared with the control group, siRNA interference
After ZNF24 gene expression, the transfer ability of cell weakens.
The above embodiments are merely illustrative of the technical solutions of the present invention, rather than is limited；Although referring to aforementioned reality
Applying example, invention is explained in detail, for those of ordinary skill in the art, still can be to aforementioned implementation
Technical solution documented by example is modified or equivalent replacement of some of the technical features；And these are modified or replace
It changes, the spirit and scope for claimed technical solution of the invention that it does not separate the essence of the corresponding technical solution.
Sequence table
<110>Shandong Inst. of Parasitic Diseases Preventing and Control
<120>application in a kind of siRNA and its inhibition cell Proliferation and migration for interfering ZNF24 gene expression
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
gcgaagaggg atcaagtatt t 21
<210> 2
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
atacttgatc cctcttcgct t 21
<210> 3
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
gcagtttgtt gccatcctat t 21
<210> 4
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
taggatggca acaaactgct t 21
<210> 5
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 5
ggatttggag agtgaacttt t 21
<210> 6
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 6
aagttcactc tccaaatcct t 21
Claims (9)
1. a kind of for interfering the ZNF24-siRNA-292 of ZNF24 gene expression, which is characterized in that described is used to interfere
The nucleotide sequence of the ZNF24-siRNA-292 of ZNF24 gene expression are as follows:
Positive-sense strand: 5 '-GCGAAGAGGGATCAAGTATTT-3 ',
Antisense strand: 5 '-ATACTTGATCCCTCTTCGCTT-3 '.
2. a kind of for interfering the ZNF24-siRNA-485 of ZNF24 gene expression, described is used to interfere ZNF24 gene expression
ZNF24-siRNA-485 nucleotide sequence are as follows:
Positive-sense strand: 5 '-GCAGTTTGTTGCCATCCTATT-3 ',
Antisense strand: 5 '-TAGGATGGCAACAAACTGCTT-3 '.
3. a kind of for interfering the ZNF24-siRNA-575 of ZNF24 gene expression, described is used to interfere ZNF24 gene expression
ZNF24-siRNA-575 nucleotide sequence are as follows:
Positive-sense strand: 5 '-GGATTTGGAGAGTGAACTTTT-3 ',
Antisense strand: 5 '-AAGTTCACTCTCCAAATCCTT-3 '.
4. of any of claims 1-3 for interfering the siRNA of ZNF24 gene expression being used to prepare inhibition cell
Application in proliferation and the preparation of migration.
5. application according to claim 4, which is characterized in that including following operating procedure:
(1) it is cracked after transfecting described ZNF24-siRNA-292, ZNF24-siRNA-485 or ZNF24-siRNA-575 into cell
Obtain total serum IgE；
(2) the first chain cDNA is synthesized by template reverse transcription of the total serum IgE in step (1)；
(3) RT-PCR is carried out as template using the cDNA in step (2) and detects the inhibition efficiency that ZNF24-siRNA expresses ZNF24；
(4) the highest ZNF24-siRNA of inhibition efficiency in step (3) is transfected into cell, preparation inhibits cell Proliferation and moves
The preparation of shifting.
6. application according to claim 5, which is characterized in that the concentration of ZNF24-siRNA is 20 in the step (1)
pmol。
7. application according to claim 5, which is characterized in that the dosage of total serum IgE is 10 ng-2 μ in the step (2)
g。
8. application according to claim 5, which is characterized in that RT-PCR reaction system is 20 μ l in the step (3),
It include: cDNA template 20-100 ng, 1 μ l of forward primer, 1 μ l, SuperReal PreMix Plus of reverse primer, 10 μ l,
RNase-free ddH2O is supplied to 20 μ l.
9. application according to claim 5, which is characterized in that RT-PCR response procedures in the step (3) are as follows: 95 DEG C pre-
It is denaturalized 15 min；95 DEG C of denaturation 10s, 60 DEG C of annealing 30s, 72 DEG C of extension 30s, totally 40 recycle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910602849.9A CN110305865A (en) | 2019-07-05 | 2019-07-05 | A kind of siRNA and its inhibition cell Proliferation for interfering ZNF24 gene expression and the application in migration |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910602849.9A CN110305865A (en) | 2019-07-05 | 2019-07-05 | A kind of siRNA and its inhibition cell Proliferation for interfering ZNF24 gene expression and the application in migration |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110305865A true CN110305865A (en) | 2019-10-08 |
Family
ID=68079282
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910602849.9A Pending CN110305865A (en) | 2019-07-05 | 2019-07-05 | A kind of siRNA and its inhibition cell Proliferation for interfering ZNF24 gene expression and the application in migration |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110305865A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101892236A (en) * | 2010-07-16 | 2010-11-24 | 武汉大学 | Construction and application of RNA interference expression vector of targeted ZNF268 gene |
| CN109806401A (en) * | 2017-11-20 | 2019-05-28 | 北京蛋白质组研究中心 | Inhibit application of the substance of ZNF8 expressing quantity in the product that preparation prevents and treats cancer |
-
2019
- 2019-07-05 CN CN201910602849.9A patent/CN110305865A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101892236A (en) * | 2010-07-16 | 2010-11-24 | 武汉大学 | Construction and application of RNA interference expression vector of targeted ZNF268 gene |
| CN109806401A (en) * | 2017-11-20 | 2019-05-28 | 北京蛋白质组研究中心 | Inhibit application of the substance of ZNF8 expressing quantity in the product that preparation prevents and treats cancer |
Non-Patent Citations (3)
| Title |
|---|
| DI JIA等: "The endogenous zinc finger transcription factor, ZNF24,modulates the angiogenic potential of human microvascular endothelial cells", 《THE FASEB JOURNAL》 * |
| JIANZHONG LI等: "A transcript profiling approach reveals the zinc finger transcription factor ZNF191 is a pleiotropic factor", 《BMC GENOMICS》 * |
| ZIYU LE等: "Predictive single nucleotide polymorphism markers for acute oral mucositis in patients with nasopharyngeal carcinoma treated with radiotherapy", 《ONCOTARGET》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108546702A (en) | The siRNA for targeting long-chain non-coding RNA DDX11-AS1 and its application in liver cancer treatment | |
| CN106995858A (en) | A kind of lncRNA related to liver cancer diagnosis and treatment | |
| CN106282347A (en) | HoxC11 as biomarker preparation adenocarcinoma of lung pre-diagnostic reagent in application | |
| CN110123828A (en) | Application of the inhibitor of PRALR in the drug that resistance to taxol oophoroma is treated in preparation | |
| CN114432452A (en) | Application of RNA Hsa _ circ _0063865 inhibitor in preparation of anti-esophageal squamous cell carcinoma drug | |
| CN110229901A (en) | Gene hsa_circ_0027089 relevant to triple negative breast cancer diagnosis and treatment and its application | |
| CN108913695B (en) | Application of ZEB1 in human heart fibroblast | |
| CN107267616A (en) | A kind of application of Noncoding gene biomarker in liver cancer | |
| CN107893115A (en) | The purposes of ALKBH1 genes and its expression product in the kit for diagnosing tumour is prepared, treat the medicine of tumour | |
| CN110305865A (en) | A kind of siRNA and its inhibition cell Proliferation for interfering ZNF24 gene expression and the application in migration | |
| CN104995300B (en) | The adjusting of RNA activity and vasopermeability | |
| CN110229900A (en) | Gene hsa_circ_0103520 relevant to breast cancer diagnosis and treatment and its application | |
| CN110317878A (en) | A kind of long-chain non-coding RNA and its application for bladder cancer diagnosis and treatment monitoring | |
| CN114457158A (en) | Application of Hsa _ circ _0006867 serving as esophageal cancer molecular target in preparation of drugs and kits | |
| CN113528523B (en) | CRRNA (crribonucleic acid) of specific targeting F3-T3 fusion gene based on CRISPR (clustered regularly interspaced short palindromic repeats) -Cas13a system and application of CRRNA | |
| CN107227362A (en) | A kind of gene related to liver cancer and its application | |
| CN110129319A (en) | A kind of siRNA of PRALR and application thereof | |
| CN105624159B (en) | A kind of siRNA and its application for people's EDIL3 gene | |
| CN110951880B (en) | Application of reagent for detecting lncRNA marker of hypopharynx cancer in preparation of product for diagnosing hypopharynx cancer | |
| CN110305864A (en) | A kind of siRNA for interfering DNALI1 gene expression and its application in inhibiting cell Proliferation and migrating | |
| CN106729756A (en) | Application of the biomarker as target in adenocarcinoma of lung diagnosis and treatment | |
| CN110257522A (en) | Gene hsa_circ_0045881 relevant to breast cancer diagnosis and treatment and its application | |
| CN107893119A (en) | Applications of the ZCCHC12 in osteosarcoma | |
| CN107881240A (en) | The diagnosis and treatment mark of osteosarcoma | |
| CN112941183B (en) | Application of non-coding gene miR-187-5p in primary liver cancer diagnosis and treatment |
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 |