CN103497953A - ShRNA (short hairpin ribonucleic acid) capable of inhibiting expression of buffalo SUV39H1 gene, lentiviral expression vector, construction method of lentiviral expression vector and applications of shRNA - Google Patents
ShRNA (short hairpin ribonucleic acid) capable of inhibiting expression of buffalo SUV39H1 gene, lentiviral expression vector, construction method of lentiviral expression vector and applications of shRNA Download PDFInfo
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Abstract
The invention discloses a shRNA (short hairpin ribonucleic acid) capable of inhibiting the expression of a buffalo SUV39H1 gene. The shRNA is shSUV39H1-259 or shSUV39H1-597 which comprises siRNA (small interfering ribonucleic acid) sense strand of 19nt, loop ring of 9nt, siRNA antisense strand of 19nt, and a termination signal. The shRNA can efficiently inhibit the expression of the buffalo SUV39H1 gene. The invention further comprises a lentiviral expression vector of the shRNA and a construction method of the lentiviral expression vector. Proved by experiments, the expression of the buffalo SUV39H1 gene can be effectively and stably inhibited by applying the shRNA, and in gene function research and the transgenic animal preparation processes, the influence and action mechanism on the expression efficiency of a foreign target transgene can be researched through inhibiting the expression of the SUV39H1 gene, thus providing a new conception and method for the search for improving the expression efficiency of the foreign gene in the transgenic animal.
Description
Technical field
The present invention relates to buffalo piebald supressor homologue 1(SUV39H1) gene, especially a kind of shRNA, Lentiviral and construction process thereof and application that suppresses buffalo SUV39H1 genetic expression.
Background technology
Buffalo is one of main large domestic animal of south China, has the biological characteristicses such as high-temp resisting high-humidity resisting, disease resistance are strong, crude feed tolerance, is applicable to very much the south China rural area and raises.The quality better of buffalo milk, be of high nutritive value, the content of the content of its dry-matter, milk fat content and total protein is 2 times of ordinary milk, particularly the content of milk casein (4.14%), be 16 times (0.26%) of Fresian, is the high quality raw material of manufacturing high-grade cheese.Therefore, develop buffalo poultry kind of a resource, by existing labour with buffalo be converted into breast with or the commodity buffalo of Dual-purpose production of milk and meat, be the trend of buffalo herding industry development.Yet the natural propagation rate of buffalo is very low, the population production performance is low, the urgent need development and improvement is a collection of improves buffalo reproductivity with the closely-related agriculture hi-tech of domestic animal genetic improvement.Utilize embryo biotechnology and clone technology, particularly the transgene clone technology, be an effective technology measure of cultivating prolificacy and the high water gaging ox new variety of giving milk.
In the transgenic animal preparation process, the expression level of foreign gene is subject to the impact of several factors, and wherein epigenetic modification is an important factor that affects exogenous gene expression efficiency.Histone methylated is one of histone modification mode, in apparent modification widely, histone methylated to chromatinic structure and function all tool play a very important role.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of shRNA, Lentiviral and construction process thereof and application that suppresses buffalo SUV39H1 genetic expression.
Adopt following technical scheme for solving the problems of the technologies described above the present invention: the shRNA that suppresses buffalo SUV39H1 genetic expression, this shRNA is shSUV39H1-259 or shSUV39H1-597, they all comprise the siRNA positive-sense strand of 19nt, the loop ring of 9nt, siRNA antisense strand and the termination signal of 19nt; The base sequence of loop ring is TTCAAGAGA; The base sequence of termination signal is TTTTTTT; The positive-sense strand of shSUV39H1-259 has the base sequence of sequence table SEQ .ID.No.1, and its antisense strand has the base sequence of sequence table SEQ .ID.No.2; The positive-sense strand of shSUV39H1-597 has the base sequence of sequence table SEQ .ID.No.3, and its antisense strand has the base sequence of sequence table SEQ .ID.No.4.
The Lentiviral of the shRNA of above-mentioned inhibition buffalo SUV39H1 genetic expression, this carrier is:
PSicoR-GFP-shSUV39H1-259 or pSicoR-GFP-shSUV39H1-597.
The construction process of the Lentiviral of the shRNA of above-mentioned inhibition buffalo SUV39H1 genetic expression, Lentiviral is cloned into respectively in Lentiviral pSicoR-GFP by synthetic shSUV39H1-259 or shSUV39H1-597 and obtains.
The application of the shRNA of above-mentioned inhibition buffalo SUV39H1 genetic expression aspect inhibition buffalo SUV39H1 genetic expression.
The application of the Lentiviral of the shRNA of above-mentioned inhibition buffalo SUV39H1 genetic expression aspect inhibition buffalo SUV39H1 genetic expression.
Application aspect the expression efficiency of the shRNA of above-mentioned inhibition buffalo SUV39H1 genetic expression foreign gene in the regulation and control transgenic cell.
Application aspect the expression efficiency of the shRNA Lentiviral of above-mentioned inhibition buffalo SUV39H1 genetic expression foreign gene in the regulation and control transgenic cell.
SUV39H1 albumen is the first histone lysine methylation transferring enzyme be identified in Mammals, a kind of single-minded the 9th Methionin of histone H 3 that acts on, it is attached to histone H 3 K9me1/2 site specifically, in epigenetic modification, the trimethylammonium of the dimethyl in the 9th the lysine residue site of histone H 3 of SUV39H1 gene institute catalysis and the particularly outstanding and main catalysis H3K9 of trimethylammonium (H3K9me2/3) effect, promote heterochromatic formation and regulatory gene to transcribe, directly affect exogenous gene expression efficiency.Except the transcriptional expression that participates in gene, the SUV39H1 gene is also relevant with apparent modifications such as DNA methylation, acetylation of histone.Therefore, be necessary for further understanding SUV39H1 and laying the foundation aspect regulating expression of foreign genes mechanism, and provide reference frame for studying its application in transgenic animal.
For this reason; the contriver utilizes molecule and cytobiology technology; at first the clone obtains buffalo piebald supressor homologue 1(SUV39H1) coding region; built buffalo SUV39H1 gene fusion protein expression vector, design has been synthesized buffalo SUV39H1 gene shRNA fragment and has built its Lentiviral.Then, adopt the liposome transfection method that buffalo SUV39H1 gene fusion protein expressing plasmid and its shRNA Lentiviral corotation are entered to the 293T cell, collecting cell detects the expression of buffalo SUV39H1 gene in the 293T cell, rapid screening obtains the efficient shRNA interference sequence that suppresses buffalo SUV39H1 gene, and attempts the application aspect the expression efficiency of its foreign gene in the regulation and control transgenic cell.Research work of the present invention comprises following basic step:
<1 > structure of the clone of buffalo SUV39H1 gene, fusion expression vector and detection thereof
The clone obtains the ORF full length sequence of buffalo SUV39H1 gene mRNA, after the sequence verification sequence is correct, by its directed insertion in the pEGFP-N1 carrier, build the fusion protein expression vector of buffalo SUV39H1 gene, enzyme is cut the positive recombinant plasmid pEGFP-N1-SUV39H1 of checking.Recombinant plasmid imports the 293T cell through the liposome transfection method, and after cell cultures 48h, observation of cell green fluorescence expression under fluorescent microscope, identify that it can be at the 293T cells.
<2 > structure, the cell detection of the design of shRNA fragment and Lentiviral thereof
For buffalo SUV39H1 gene C Ds sequence, with reference to the siRNA principle of design, select 2 siRNA sequences as target site (259bp-278bp, 597bp-617bp), the BLAST comparison confirms that above sequence and other gene orders are without homology.It is designed to the shRNA structure, the siRNA positive-sense strand that the shRNA structure comprises 19nt, the loop ring (TTCAAGAGA) of 9nt, the siRNA antisense strand of 19nt and termination signal (TTTTTTT), restriction enzyme site XhoI, NotI are introduced in the shRNA two ends.Difference called after shSUV39H1-259, shSUV39H1-597.Select an irrelevant sequence shRNA1864 (N.C) as negative control simultaneously.Synthetic shRNA sequence also is cloned in Lentiviral pSicoR-GFP, and enzyme is cut and sequence verification.ShRNA Lentiviral (pSicoR-GFP-shSUV39H1-259/597/N.C) imports 293T cell, 48h after transfection, each shRNA fragment expression situation on the observation of cell level under fluorescent microscope through the liposome transfection method.
<3 > suppress the screening of buffalo SUV39H1 genetic expression shRNA sequence
Adopt the method for cotransfection, goal gene buffalo SUV39H1 expression vector (target plasmid) and shRNA expression vector (interference plasmid) are entered to the 293T cell with the 1:3 mass ratio through the liposome cotransfection, after transfection 48h, observation of cell fluorescence situation under fluorescent microscope.Expression by real-time quantitative PCR (Real-Time RT-PCR) method at mRNA level detection SUV39H1 gene, calculate each shRNA fragment suppression efficiency.
By above-mentioned steps, the contriver has obtained shRNA sequence shSUV39H1-259 and the shSUV39H1-597 that efficient stable of the present invention suppresses buffalo SUV39H1 genetic expression fast.Experiment shows, when target plasmid (pEGFP-N1-SUV39H1) and interference plasmid (pSicoR-GFP-shSUV39H1-259/597/N.C) during with the transfection of 1:3 ratio, shSUV39H1-259 and shSUV39H1-597 to the suppression efficiency of SUV39H1 be respectively 33.85% and 78.75%, shSUV39H1-597 suppression efficiency higher than shSUV39H1-259.As previously mentioned, in conjunction with the present invention, in gene functional research and transgenic animal preparation process, can inquire into it by inhibition SUV39H1 genetic expression and external source be turned to impact and the mechanism of action thereof of destination gene expression efficiency, for finding the expression efficiency that improves foreign gene in transgenic animal, provide new thinking and method.
The accompanying drawing explanation
Fig. 1 is the structural representation of buffalo SUV39H1 gene fusion protein expression vector pEGFP-N1-SUV39H1.
Fig. 2 is the structural representation of the Lentiviral pSicoR-GFP-shSUV39H1-259/597/N.C of shRNA.
Fig. 3 is Fluirescence observation figure as a result after target plasmid and interference plasmid (mass ratio 1:3) cotransfection 293T cell 48h, in figure: A:pEGFP-N1-SUV39H1 plasmid and pSicoR-GFP-1864 plasmid, B:pEGFP-N1-SUV39H1 plasmid and pSicoR-GFP-SUV39H1shRNA259 plasmid, C:pEGFP-N1-SUV39H1 plasmid and pSicoR-GFP-SUV39H1shRNA597 plasmid, the D:pEGFP-N1-SUV39H1 plasmid, E: plasmid-free contrast.
Fig. 4 is that target plasmid and interference plasmid (mass ratio 1:3) cotransfection 293T cell real-time fluorescence quantitative PCR detect figure (48h), in figure: SUV39H1 is SUV39H1 target plasmid, the 1864th, contrast 1864shRNA, shRNA1 is shSUV39H1-259, shRNA2 is shSUV39H1-597, wherein difference goes up the different remarkable a of target value difference, b, c (P<0.05).
Fig. 5 is that target plasmid and interference plasmid (mass ratio 1:3) cotransfection 293T cell Western-blot detect figure (72h), in figure: A figure is with β-actin antibody, B figure is with SUV39H1 antibody, 1:SUV39H1 target plasmid, 2: contrast shRNA1864,3:shSUV39H1-259,4:shSUV39H1-597.
Fig. 6 is that SUV39H1 gene shRNA virus infection turns IFN α-2b gene Bcap-37 clone sample SUV39H1 gene qRT-PCR detected result figure, the different upper different remarkable a of target value difference in figure, b, c, d (P<0.05)
Fig. 7 is that SUV39H1 gene shRNA virus infection turns IFN α-2b gene Bcap-37 clone sample external source IFN α-2b gene qRT-PCR detected result figure, the different upper different remarkable a of target value difference in figure, b, c, d (P<0.05)
Embodiment
One, the structure of correlative expression vector
1. the structure of buffalo SUV39H1 expression vector
Ox SUV39H1 gene mRNA [NM_001046264.2] primers of announcing according to NCBI GenBank, and introduce respectively XhoI, EcoRI restriction enzyme site at 5 ' end of upstream and downstream primer, primer is:
SUV39H1-upstream 5 '-CTCGAGGGAAAGATGGCGGAAAAT-3 ' (sequence table SEQ .ID.No.7),
SUV39H1-downstream 5 '-GAATTCCCGAAGAGGTATTTGCGGCAG-3 ' (sequence table SEQ .ID.No.8).
From slaughterhouse water intaking ox fresh ovarian tissues, put into and be added with in advance the protectant 2ml EP of 1ml RNA pipe.Extract total RNA with the Trizol cracking process, reverse transcription is cDNA.Take buffalo ovaries cDNA as template, amplification obtains the ORF full length sequence (sequence table SEQ .ID.No.9) of buffalo SUV39H1 gene, length is about 1100bp, after being reclaimed, the PCR product inserts the pMD18-T carrier, called after pMD18-T-SUV39H1, cut evaluation with EcoRI and XhoI enzyme, enzyme is cut product can see that through agarose gel electrophoresis length is respectively the band of 2.8Kb and 1.1Kb, proof is inserted, recombinant plasmid pMD18-T-SUV39H1 is served to the Hai Shenggong order-checking, and recording sequence length is 1128bp.Sequencing result is compared through Blast, with ox SUV39H1 DNA homolog in NCBI be 99%.
With XhoI and EcoRI double digestion pMD18-T-SUV39H1, glue reclaims the fragment of 1100bp.With same enzyme linearizing pEGFP-N1 carrier.By in the directed insertion vector pEGFP-N1 of SUV39H1 gene fragment, recombinant plasmid carries out enzyme with XhoI and EcoRI and cuts evaluation correctly.Buffalo SUV39H1 fusion protein expression vector pEGFP-N1-SUV39H1 structure as shown in Figure 1.
2.shRNA the structure of Lentiviral
Adopt the siRNA design software, for SUV39H1 gene ORF sequence, select 2 different target sites as target sequence (259bp-278bp, 597bp-617bp), and the shRNA structure of the siRNA antisense strand of the loop ring (TTCAAGAGA) of the siRNA positive-sense strand that it is designed to comprise 19nt, 9nt and 19nt, distinguish called after shSUV39H1-259 and shSUV39H1-597; Select an irrelevant sequence shRNA-1864 (N.C) as negative control (in Table 1) simultaneously.Synthetic 2 shRNA sequences and 1 negative control sequence, and be cloned in Lentiviral pSicoR-GFP, sequence verification, and verify with XhoI and NotI double digestion.The shRNA expression vector structure built as shown in Figure 2.
Table 1shRNA sequential structure
Two, expression of recombinant plasmid detects
1. the cell expressing of recombinant plasmid pEGFP-N1-SUV39H1 detects
The pEGFP-N1-SUV39H1 recombinant plasmid of structure is used to extract and be diluted to final concentration without the intracellular toxin plasmid extraction kit be 1 μ g/ μ L, with LTX liposome transfection 293T cell.Transfection plasmid amount is 2.5 μ g, does untransfected group and empty carrier (pEGFP-N1) transfection group in contrast simultaneously.After transfection 48h, under fluorescent microscope, observe, recombinant vectors transfection group and empty carrier transfection group all can observe the expression of green fluorescent protein EGPF.Show that constructed buffalo SUV39H1 gene fusion protein expression vector (pEGFP-N1-SUV39H1) can be at 293T cell transient expression.
2. the detection of expression of recombinant plasmid pSicoR-GFP-shSUV39H1-259/597/N.C
With reference to step (two, 1), the shRNA Lentiviral is transfected into the 293T cell through liposome method.Can observe the expression of green fluorescence EGFP after 48h under fluorescent microscope.
Three, target plasmid and interference plasmid cotransfection 293T cell
By cell with 2 * 10
6density is inoculated in 6 porocyte culture plates, carries out transfection when cell degree of converging reaches 80%.It is 4 groups that test is divided into, and is respectively untransfected group, goal gene transfection group, target gene and interference plasmid cotransfection group (shSUV39H1-259, shSUV39H1-597) and negative control group (shRNA1864-N.C).According to Lipofectamine2000 specification sheets mixing plasmid and Lipofectamine2000, both ratios are 10 μ l:4 μ g(1 μ g SUV39H1 and 3 μ g shSUV39H1), plasmid-Lipofectamine2000 mixture is splashed into and treats the transfectional cell hole.Be replaced by after 6h and contain 10% serum, 1% pair of anti-DMEM nutrient solution, observation transfection effect under fluorescent microscope after 48h.The cotransfection group can be seen respectively the expression (seeing Fig. 3) of reporter gene green fluorescent protein.
Four, real-time quantitative PCR and Western-blot method detect the suppression efficiency of shRNA to the SUV39H1 gene
In step (two), target plasmid and interference plasmid with mass ratio 1:3 cotransfection 293T cell 48h after, collecting cell.In each sample, add 1ml Trizol to extract cell total rna.After DNaseI fully digests DNA residual in total RNA, with the AMV reversed transcriptive enzyme, by the RNA reverse transcription, be cDNA.Reaction conditions is: 25 ℃ of 5min, 42 ℃ of 60min, 95 ℃ of 5min.Measure the OD value of reverse transcription product cDNA, use RNase-Free ddH
2o dilution template final concentration is 100ng/ μ L, for real-time quantitative RT-PCR, detects.The QRT-PCR primer is as follows:
SUV39H1-Fr5 '-CGAGCCGGGCTGCCCATCTA-3 ' (sequence table SEQ .ID.No.10),
SUV39H1-Rv5 '-CCCAGCCGCGTCCATCATCC-3 ' (sequence table SEQ .ID.No.11),
β-actin-Fr5 '-GCCCTGGCACCCAGCACAAT-3 ' (sequence table SEQ .ID.No.12),
β-actin-Rv5 '-GGAGGGGCCGGACTCATCGT-3 ' (sequence table SEQ .ID.No.13).
The real-time quantitative PCR amplification condition is 50 ℃ of 2min, 95 ℃ of 10min, 95 ℃ 15s30 circulation, 60 ℃ of 1min.According to 2
-△ △ Ctmethod is calculated the relative expression quantity of buffalo SUV39H1 gene.When target plasmid and interference plasmid, during with 1:3 ratio cotransfection 293T cell, shSUV39H1-259 and shSUV39H1-597 are respectively 33.85% and 78.75% to the suppression efficiency of SUV39H1.ShSUV39H1-259 and shSUV39H1-597 fragment all have interference effect, wherein the higher (see figure 4) of the inhibition of shSUV39H1-597.
Carry out equivalent loading, low high-voltage leakage of electricity swimming, transferring film, sealing, antibody incubation, gel imaging according to the Western-blot routine operation.As shown in Figure 5, β between each group-actin protein band is more clear consistent; In feminine gender and blank group, SUV39H1 protein expression band is clear, and in shRNAs transfection experiment group, the SUV39H1 protein band all obviously weakens.
Five, SUV39H1 gene shRNAs is the impact of related gene expression on turning IFN α-2b gene cell.
Adopt calcium phosphate transfection method packing SUV39H1 gene shSUV39H1-259 and the high titre slow virus of shSUV39H1-597.Utilize the Bcap-37 clone of virus infection stably express IFN α-2b gene, collecting cell sample after 72h, adopt the qRT-PCR method to detect SUV39H1, IFN α-2b genetic expression.Result shows, under identical virus quantity infectious condition, a SUV39H1 gene shRNA-259 and shRNA-5972 fragment all has significant inhibition to SUV39H1 genetic expression, jamming effectiveness be respectively 53.07% and 31.28%(Fig. 6).With negative control group, compare, in 2 shRNAs virus infected cells, the expression amount of IFN α-2b gene significantly improves (P<0.05), improve respectively 96.25% and 121.08%(Fig. 7).Result shows that the Bcap-37 clone of SUV39H1 gene shRNA slow virus infection expression IFN2b-α gene can improve the wherein expression level of external source IFN2b-α, illustrates that regulation and control SUV39H1 genetic expression can be used for regulating the genetically modified expression efficiency of external source.
Claims (7)
1. a shRNA who suppresses buffalo SUV39H1 genetic expression, it is characterized in that: this shRNA is shSUV39H1-259 or shSUV39H1-597, they all comprise the siRNA positive-sense strand of 19nt, the loop ring of 9nt, siRNA antisense strand and the termination signal of 19nt;
The base sequence of described loop ring is TTCAAGAGA;
The base sequence of described termination signal is TTTTTTT;
The positive-sense strand of described shSUV39H1-259 has the base sequence of sequence table SEQ .ID.No.1, and its antisense strand has the base sequence of sequence table SEQ .ID.No.2;
The positive-sense strand of described shSUV39H1-597 has the base sequence of sequence table SEQ .ID.No.3, and its antisense strand has the base sequence of sequence table SEQ .ID.No.4.
2. the Lentiviral that suppresses according to claim 1 the shRNA of buffalo SUV39H1 genetic expression is characterized in that this carrier is:
PSicoR-GFP-shSUV39H1-259 or pSicoR-GFP-shSUV39H1-597.
3. suppress according to claim 2 the construction process of Lentiviral of the shRNA of buffalo SUV39H1 genetic expression, it is characterized in that: described Lentiviral is cloned into respectively in Lentiviral pSicoR-GFP by synthetic described shSUV39H1-259 or shSUV39H1-597 and obtains.
4. suppress according to claim 1 the application of shRNA aspect inhibition buffalo SUV39H1 genetic expression of buffalo SUV39H1 genetic expression.
5. suppress according to claim 2 the application of Lentiviral aspect inhibition buffalo SUV39H1 genetic expression of the shRNA of buffalo SUV39H1 genetic expression.
6. the application aspect the expression efficiency of the shRNA that suppresses according to claim 1 buffalo SUV39H1 genetic expression foreign gene in the regulation and control transgenic cell.
7. the application aspect the expression efficiency of the shRNA Lentiviral that suppresses according to claim 2 buffalo SUV39H1 genetic expression foreign gene in the regulation and control transgenic cell.
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| CN109628451A (en) * | 2019-01-10 | 2019-04-16 | 广西大学 | It is a kind of inhibit rabbit Deptor gene expression shRNA, Lentiviral and its construction method and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174577A (en) * | 2011-01-24 | 2011-09-07 | 广西大学 | Method for producing genetically modified water buffalos by utilizing lentiviral vector |
| CN102517287A (en) * | 2012-01-10 | 2012-06-27 | 广西大学 | Short hairpin ribonucleic acid (shRNA) for inhibiting expression of buffalo triggering receptor expressed on myeloid cells 1 (TREM1) gene, lentiviral expression vector of shRNA and construction method and application of lentiviral expression vector |
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| CN102174577A (en) * | 2011-01-24 | 2011-09-07 | 广西大学 | Method for producing genetically modified water buffalos by utilizing lentiviral vector |
| CN102517287A (en) * | 2012-01-10 | 2012-06-27 | 广西大学 | Short hairpin ribonucleic acid (shRNA) for inhibiting expression of buffalo triggering receptor expressed on myeloid cells 1 (TREM1) gene, lentiviral expression vector of shRNA and construction method and application of lentiviral expression vector |
Non-Patent Citations (2)
| Title |
|---|
| A LAKSHMIKUTTYAMMA ET AL.: "Reexpression of epigenetically silenced AML tumor suppressor genes by SUV39H1 inhibition.", 《ONCOGENE》 * |
| GENBANK: "NM_001046264", 《GENBANK》 * |
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|---|---|---|---|---|
| CN109628451A (en) * | 2019-01-10 | 2019-04-16 | 广西大学 | It is a kind of inhibit rabbit Deptor gene expression shRNA, Lentiviral and its construction method and application |
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