CN104830903A - Polypeptide K12-based transgenic vector and application thereof - Google Patents
Polypeptide K12-based transgenic vector and application thereof Download PDFInfo
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Abstract
The invention relates to a polypeptide K12 and a K12-modified gene vector. The sequence of the polypeptide K12 is Lys-Lys-Lys-Arg-Lys-Cys-Gly-Asn-Lys-Arg-Thr-Arg; and the gene vector is prepared by the following steps: firstly, selecting poloxamer 407 (P407) and connecting to PEI (polyether-imide) to obtain a high molecular weight PEI derivative with a multi-branch form or a network structure; selecting a tLyP-1 oligopeptide, connecting with a nuclear localization signal peptide (NLS), and synthesizing the polypeptide K12 which targets NRP and is capable of improving the nuclear delivery ability; coupling the K12 to the EPI derivative by employing a crosslinking technique, and building a novel non-viral gene vector P407-PEI-K12. The cytotoxicity and transfection experiment results prove that the novel non-viral gene vector system P407-PEI-K12 provided by the invention is low in toxicity and has relatively high targeting property; and the cell and in-vivo transfection effects of the gene vector are superior to those of a control group.
Description
Technical field
The present invention relates to medical art, specifically, is PEI derivative P407-PEI transgene carrier and the application thereof of polypeptide K12 modification.
Background technology
Gene therapy is based upon the novel method for the treatment of in genetic engineering technique and molecular genetics principle in recent years, and the Basic of Biology betiding development because of tumour is transgenation, so gene therapy has now become the merit gram field that tumour most is wished.Gene therapy has three important steps, i.e. goal gene, transgene carrier and target cell.Gene pours the core technology that system is gene therapy into, and the biggest problem that present stage faces is not yet to find desirable genophore.The carrier of current application comprises virus vector and the large class of non-virus carrier two.Virus vector transfection efficiency higher but exist carrying capacity low, have the problems such as potential security threat, and non-virus carrier development in recent years is rapid, especially cationic polymers.
Polymine (polyethylenimine PEI) is Recent study cationic polymer non-viral gene vector the most widely, is rich in positively charged ion, has powerful surge capability, has the ability more by force in conjunction with DNA and adherent cell.But polymine uses existence two outstanding problems as genophore: the first, transfection efficiency and cytotoxicity exist contradiction.Though small molecules PEI cytotoxicity is low, easily dissociate with DNA under physiological ionic concn, cause transfection poor; Though molecular weight has comparatively ideal transfection efficiency at the PEI of more than 20kd, because PEI rich surface is containing non-biodegradable in positive charge and body, high molecular PEI is caused to show stronger cytotoxicity.Second, polymine targeting is poor: it be utilize self with positive charge, the acceptor electronegative with cell surface is combined by electrostatic interaction, so the selection poor specificity of cell, solving targeting problem has become the problem paid close attention to the most in non-virus carrier.In addition, polymkeric substance/DNA mixture needs to enter into nucleus, and dissociated by RNA thus play a role.Therefore, how to improve cationic compound in vivo the nucleus delivery capability of target cell become it and can be applied to one of clinical key.Poloxamer poloxame is the PEO-PPO-PEO type nonionic triblock copolymer that a class is made up of polyoxyethylene (PEO), polyoxypropylene (PPO).Poloxamer188 (be called for short P407) is made up of about 70% ethylene oxide and 30% propylene oxide, molecular-weight average 11500, be study in Families of poloxamers the most extensively, be also the most frequently used a kind of polymkeric substance arrived.
Nuclear localization signal NLS is that in core, functional protein enters nuclear architecture basics, that some albumen of mediation enters one section of the core fully information segment of necessity, macromolecular substance by the mediation of NLS through nuclear Pore Complex active transport in nucleus, wherein the NLS of most study is the T antigen from SV40, and major function sequence is KKKRKV.Can the complex by intracellular consideration convey of PEI be transported in core by NLS, improve the transfection efficiency of mixture.NPR is the transmembrane protein that cerebral glioma tumor cell membrane and neovascular endothelium cytolemma exist, play a part very crucial in the biology of tumour, confirm that NRP acceptor all exists high expression level, one of mark that can be used as malignant tumour on the cells such as carcinoma of the pancreas, cancer of the stomach, colorectal carcinoma, melanoma, ovarian cancer at present.TLyP-1 is newfound target polypeptide, and aminoacid sequence is CGNKRTR, and the specific target of energy is also efficiently combined with NRPs acceptor, can produce across Vascular permeability and tumour membrane penetration effect simultaneously, have specificity and high-affinity to NPR acceptor.Research display tLyP-1 can optionally go back to the nest and tumour target position, is seeped into blood vessel efficiently from Ink vessel transfusing, and homogeneity ground infiltrates in mesenchyma stroma of tumors, and can enter in karyocyte through the cytolemma of tumour cell.
Therefore, tLyP-1 (sequence is CGNKRTR) can be introduced NLS sequence (KKKRK) in theory, (sequence is KKKRKCGNKRTR to synthesize the polypeptide tLyP-1-1-NLS containing tLyP-1, NLS, called after K12), for modifying P407-PEI derivative, strengthen it in vivo to tumor cells selectivity, and improve core delivery capability, thus transfection efficiency in the body of increase DNA.
Chinese patent literature CN201110372844.5, disclose genophore and its preparation method and application that a kind of bifunctional peptide is modified, the genophore of this invention is by the polyethylenimine derivates coupling of bifunctional peptide (RGDCTAT) and pluronic P123 modification.Chinese patent literature CN201410766122.1 discloses a kind of genophore modified by functional peptides R11 and its preparation method and application, and the genophore of this invention is that the polyethylenimine derivates coupling of being modified by bifunctional peptide (RGDCNLS) and F68 P123 forms.But about poloxamer188-polymine that polypeptide tLyP-1-NLS modifies, yet there are no report.
Summary of the invention
The object of the invention is, for deficiency of the prior art, to provide a peptide species.
Of the present invention again one object be that a kind of genophore is provided.
Another object of the present invention provides the preparation method of said gene carrier.
4th object of the present invention provides a kind of gene composite.
5th object of the present invention provides the purposes of aforementioned polypeptides and genophore.
For realizing above-mentioned first object, the technical scheme that the present invention takes is:
One peptide species, described polypeptid acid sequence is as shown in SEQ ID NO.1.
Polypeptide provided by the invention by nuclear localization signal peptide NLS (KKKRK) with there is target form in the tLyP-1 of NRP transmembrane protein acceptor, peptide sequence is KKKRKCGNKRTR.
For realizing above-mentioned second object, the technical scheme that the present invention takes is:
A kind of genophore, it is formed by polypeptide as above and polyethylenimine derivates coupling, described polyethylenimine derivates is polymine and the P407-polymine of poloxamer188 modification, and described poloxamer188 and the mol ratio of polymine are 1:1-20; The molecular weight ranges of described polymine is 600-70000Da.
Described poloxamer188 and polymine mol ratio are 1:1-10.
Described poloxamer188 and polymine mol ratio are 1:10.
Described polypeptide and the mol ratio of polyethylenimine derivates are 1-20:1; Be preferably 2:1.
For realizing above-mentioned 3rd object, the technical scheme that the present invention takes is:
As above the preparation method of arbitrary described genophore, comprises the steps:
A () synthesizes polypeptide as above;
B () poloxamer188 dewaters after, add the triphosgene of twice molar weight, N-hydroxy-succinamide is added after magnetic agitation reaction, dropwise anhydrous triethylamine is added under agitation condition, question response, completely afterwards except desolventizing, is residue obtainedly dissolved in ethyl acetate, centrifuging and taking supernatant liquor, revolve and steam removing ethyl acetate, the poloxamer188 that must activate after cooling curing;
C poloxamer188 that () activates and polymine PEI are dissolved in anhydrous methylene chloride respectively, are added in liquid at the bottom of anhydrous methylene chloride by two liquid of gained simultaneously, the saturated rear magnetic stirrer over night of inflated with nitrogen, dialysis, and freeze-drying, obtains P407-PEI;
(d) by surfactant dissolves in dimethyl sulphoxide solution, polypeptide prepared by step (a) and the P407-PEI prepared by step (c) are dissolved in PBS solution respectively, surfactant soln is joined the P407-PEI liquid obtaining maleimation in P407-PEI liquid, the polypeptide solution prepared is joined in the P407-PEI liquid of maleimation, 4 DEG C of shake reactions are spent the night, centrifugal freeze-drying, both.
Described tensio-active agent is SMCC, is the bifunctional coupling agent that a class contains N-hydroxy-succinamide (NHS) active ester and maleimide.
First the present invention selects poloxamer188 to connect PEI, obtain multi-branched shape or cancellated high molecular PEI derivative, then utilize transmembrane protein acceptor NRP in the feature of human tumor cells and tumor neogenetic blood vessels high expression level, select the tLyP-1 small peptide of special affine this receptor, be connected with nuclear localization signal peptide NLS, synthesis has target in NRP and the polypeptide K12 improving core delivery capability, utilize crosslinking technological by the coupling of K12 and PEI derivative, thus build novel non-viral gene vector system P407-PEI-K12.
For realizing above-mentioned 4th object, the technical scheme that the present invention takes is:
As above the mixture that formed of arbitrary described genophore and DNA.
Described DNA is containing reporter gene, antioncogene and/or cytokine gene.
For realizing above-mentioned 5th object, the technical scheme that the present invention takes is:
The application in the medicine of preparation gene therapy of polypeptide as above and genophore.
The invention has the advantages that:
First the present invention selects poloxamer188 to connect PEI, obtain multi-branched shape or cancellated high molecular PEI derivative, then utilize transmembrane protein acceptor NRP in the feature of human tumor cells and tumor neogenetic blood vessels high expression level, select the tLyP-1 small peptide of special affine this receptor, be connected with nuclear localization signal peptide NLS, synthesis has target in NRP and the polypeptide K12 improving core delivery capability, utilize crosslinking technological by the coupling of K12 and PEI derivative, thus build novel non-viral gene vector system P407-PEI-K12.Cytotoxicity and transfection experiment result show, not only toxicity is low for novel non-viral gene vector system P407-PEI-K12 provided by the invention, and has stronger targeting, and in cell and body, transfection is all better than control group.
Accompanying drawing explanation
Accompanying drawing 1 is P407-PEI-K12 cytotoxicity experiment result.Wherein, 3 pillars of first bunch represent P123-PEI-K12, PEI 2KDa, PEI 25KDa from left to right respectively from left to right.
Accompanying drawing 2 is P407-PEI-K12 in-vitro transfection experimental results.Wherein, 3 pillars of first bunch represent P123-PEI-K12, PEI 25KDa, PEI 2KDa from left to right respectively from left to right.
Embodiment
Below in conjunction with accompanying drawing, embodiment provided by the invention is elaborated, but embodiments of the invention are not limited only to this.
Herein, SMCC is the bifunctional coupling agent that a class contains N-hydroxy-succinamide (NHS) active ester and maleimide, can by respectively containing sulfydryl and amino compound be connected to together with, its molecular structure is as follows.
The preparation of the P407-PEI that embodiment 1 polypeptide K12 modifies and functional verification ()
1, the preparation of P407-PEI
Take the poloxamer188 (P407,0.6mmol) after dewatering in right amount, be dissolved in the mixing solutions of dry toluene and anhydrous methylene chloride, add the triphosgene of 1.2mmol, room temperature lower magnetic force stirring reaction spends the night.Vacuum is revolved and is steamed except desolventizing, then dissolves with appropriate dry toluene and anhydrous methylene chloride, then adds 2.0mmol N-hydroxy-succinamide, under magnetic agitation, is dropwise added in reaction solution by 2.0mmol anhydrous triethylamine, continues stirring reaction and is about 4h.Question response completely after, by reacting liquid filtering and again vacuum is revolved and is steamed except desolventizing, is residue obtainedly dissolved in ethyl acetate, gets supernatant liquor, rotary evaporation, flings to ethyl acetate, cooling curing reactant, the P407 after both must activating after high speed centrifugation.PEI 2K (0.20g, be dissolved in 10mL anhydrous methylene chloride after 0.10mmol) dewatering, P407 (0.01mmol) after activation is dissolved in anhydrous methylene chloride, above-mentioned two liquid are instilled in liquid at the bottom of 10mL anhydrous methylene chloride slowly simultaneously, the saturated rear room temperature magnetic stirrer over night of nitrogen, dialysis, freeze-drying, obtains P407-PEI.
2, the synthesis Lys Lys-Lys-Arg-Lys-Cys-Gly-Asn-Lys-Arg-Thr-Arg of polypeptide K12
Aminoacid sequence according to tLyP-1: Cys-Gly-Asn-Lys-Arg-Thr-Arg, the aminoacid sequence of NLS: Lys-Lys-Lys-Arg-Lys, adopt Solid phase synthesis polypeptide tLyP-1-NLS by the biochemical company limited of Shanghai gill, its aminoacid sequence is: Lys-Lys-Lys-Arg-Lys-Cys-Gly-Asn-Lys-Arg-Thr-Arg (SEQ ID NO.1) called after K12.
3, polypeptide K12 is to the modification of P407-PEI
SMCC is dissolved in DMSO solution, NLS-tLyP-1 (polypeptide K12) and P407-PEI are dissolved in PBS solution respectively, by SMCC liquid in molar ratio 2:1 dropwise add in PEI liquid, limit edged stirs, at room temperature react 30mim, remove unconjugated SMCC by gel chromatographic columns, obtain the P407-PEI liquid (maleimided P407-PEI) of maleimation.By K12 liquid in molar ratio 2:1 add in maleimidedP407-PEI liquid, 4 DEG C shake reactions spend the night.Then use ultra-filtration centrifuge tube centrifugal, freeze-drying, obtains P407-PEI-K12.
4, the cell toxicity test of P407-PEI-K12
B16 cell is inoculated on 96 orifice plates, cultivates 48h, make cell confluency degree reach 70%-80%.Before test, suck substratum, every hole adds different concns cationic polymers (4,6,8,16,24,32 μ g/mL, serum-free 1640 is as solvent), continue to cultivate 72h, mtt assay detects cytotoxicity, and as can be seen from Figure 1, the PEI 25K cytotoxicity of unmodified is stronger, and P407-PEI-K12 is almost non-toxic, ensure that the feasibility of P407-PEI-K12 as genophore.
Table 1 cell toxicity test result
5, the in-vitro transfection experiment of P407-PEI-K12
B16 cell is inoculated on 24 orifice plates, cultivates 48h, make cell confluency degree reach 70%-80%.By P407-PEI-K12 and Luci reporter gene, 5:1,10:1,20:1 and 30:1 make mixture in mass ratio, add on 24 orifice plates, cultivate 4h, replace and continue to cultivate 48h containing blood serum medium, detect luciferin expression of enzymes, as can be seen from Figure 2, P407-PEI-K12 shows very strong transfection efficiency, far away higher than control group PEI 2K, also high than the optimum expression of PEI 25K 1000 times, illustrate that P407-PEI-K12 can well carry out transfection in cell by target.
Table 2 in-vitro transfection experimental result
Transfection experiment in the body of 6, P407-PEI-K12
Select ICR experimental mouse to set up B16 murine melanoma transplantation model, and laboratory animal is divided into 2 groups: P407-PEI-K12, w/w=30; P123-PEI-R11 (formed by polypeptide R11 and the coupling of F68 P123-polymine, synthetic method is see patent 201410766122.1), w/w=30.With plasmid PGL3-Control (being purchased from Promega) for reporter gene, to evaluate after intravenously administrable 24h the organ luciferase expression situations such as mouse core, liver, spleen, lung, kidney.Wherein, w/w=30 represents that the mass ratio of cationic polymers and Luci reporter gene is 30, and result is as shown in table 3.As can be seen from Table 3, P407-PEI-K12 shows very strong transfection efficiency, and the expression intensity of luciferase, far away higher than control group P123-PEI-R11, is 15 times of P123-PEI-R11 expression intensity at the heart, liver, spleen, lung, kidney place respectively.
Transfection efficiency experimental result in table 3 body
The preparation of the P407-PEI that embodiment 2 polypeptide K12 modifies and functional verification (two)
The preparation of P407-PEI-K12 and in-vitro transfection experimental procedure are with embodiment 1, difference is: in the present embodiment, PEI molecular weight is 70KDa, during preparation P407-PEI, P407 and PEI mol ratio used is 1:1, and the mol ratio of polypeptide K12 and P407-PEI used during synthesis P407-PEI-K12 is 5:1.In body, transfection experiment result shows: the expression intensity of P407-PEI-K12 fluorescence mycin, far away higher than P123-PEI-R11, is 12,13,12,11,11 times of P123-PEI-R11 expression intensity at the heart, liver, spleen, lung, kidney place respectively.
The preparation of the P407-PEI that embodiment 3 polypeptide K12 modifies and functional verification (three)
The preparation of P407-PEI-K12 and in-vitro transfection experimental procedure are with embodiment 1, difference is: in the present embodiment, PEI molecular weight is 4KDa, during preparation P407-PEI, P407 and PEI mol ratio used is 1:5, and the mol ratio of polypeptide K12 and P407-PEI used during synthesis P407-PEI-K12 is 1:1.In body, transfection experiment result shows: the expression intensity of P407-PEI-K12 fluorescence mycin, far away higher than P123-PEI-R11, is 13,14,11,12,12 times of P123-PEI-R11 expression intensity at the heart, liver, spleen, lung, kidney place respectively.
The preparation of the P407-PEI that embodiment 4 polypeptide K12 modifies and functional verification (four)
The preparation of P407-PEI-K12 and in-vitro transfection experimental procedure are with embodiment 1, difference is: in the present embodiment, PEI molecular weight is 600Da, during preparation P407-PEI, P407 and PEI mol ratio used is 1:20, and the mol ratio of polypeptide K12 and P407-PEI used during synthesis P407-PEI-K12 is 20:1.In body, transfection experiment result shows: the expression intensity of P407-PEI-K12 fluorescence mycin, far away higher than P123-PEI-R11, is 10,11,13,11,12 times of P123-PEI-R11 expression intensity at the heart, liver, spleen, lung, kidney place respectively.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the inventive method; can also make some improvement and supplement, these improve and supplement and also should be considered as protection scope of the present invention.
Claims (10)
1. a peptide species, is characterized in that, described polypeptid acid sequence is as shown in SEQ ID NO.1.
2. a genophore, it is characterized in that, it is formed by polypeptide according to claim 1 and polyethylenimine derivates coupling, described polyethylenimine derivates is polymine and the P407-polymine of poloxamer188 modification, and described poloxamer188 and the mol ratio of polymine are 1:1-20; The molecular weight ranges of described polymine is 600-70000Da.
3. genophore according to claim 2, is characterized in that, described poloxamer188 and polymine mol ratio are 1:1-10.
4. genophore according to claim 2, is characterized in that, described poloxamer188 and polymine mol ratio are 1:10.
5. genophore according to claim 2, is characterized in that, described polypeptide and the mol ratio of polyethylenimine derivates are 1-20:1.
6. genophore according to claim 2, is characterized in that, described polypeptide and the mol ratio of polyethylenimine derivates are 2:1.
7. the preparation method of the arbitrary described genophore of claim 2-6, is characterized in that, comprise the steps:
A () synthesizes polypeptide as claimed in claim 1;
B () poloxamer188 dewaters after, add the triphosgene of twice molar weight, N-hydroxy-succinamide is added after magnetic agitation reaction, dropwise anhydrous triethylamine is added under agitation condition, question response, completely afterwards except desolventizing, is residue obtainedly dissolved in ethyl acetate, centrifuging and taking supernatant liquor, revolve and steam removing ethyl acetate, the poloxamer188 that must activate after cooling curing;
C poloxamer188 that () activates and polymine PEI are dissolved in anhydrous methylene chloride respectively, are added in liquid at the bottom of anhydrous methylene chloride by two liquid of gained simultaneously, the saturated rear magnetic stirrer over night of inflated with nitrogen, dialysis, and freeze-drying, obtains P407-PEI;
(d) by surfactant dissolves in dimethyl sulphoxide solution, polypeptide prepared by step (a) and the P407-PEI prepared by step (c) are dissolved in PBS solution respectively, surfactant soln is joined the P407-PEI liquid obtaining maleimation in P407-PEI liquid, the polypeptide solution prepared is joined in the P407-PEI liquid of maleimation, 4 DEG C of shake reactions are spent the night, centrifugal freeze-drying, both.
8. the mixture that formed of the arbitrary described genophore of claim 2-6 and DNA.
9. mixture according to claim 8, is characterized in that, described DNA is containing reporter gene, antioncogene and/or cytokine gene.
10. polypeptide according to claim 1 and the application of the arbitrary described genophore of claim 2-6 in the medicine of preparation gene therapy.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106480092A (en) * | 2016-10-20 | 2017-03-08 | 上海海洋大学 | A kind of PEGization Fe being modified based on Functional Polypeptides R93+/ PEI genophore and its preparation method and application |
| CN106801068A (en) * | 2017-01-12 | 2017-06-06 | 西安交通大学 | A kind of non-viral gene vector of autofluorescence degradable poly citrate and preparation method thereof |
| CN109310729A (en) * | 2016-03-31 | 2019-02-05 | 慕尼黑大学 | For improving the synthesis compound of transfection efficiency |
| CN109486827A (en) * | 2018-12-04 | 2019-03-19 | 南京林业大学 | A kind of apoferritin nanocages and preparation method thereof of tumor-homing cell-penetrating peptide tLyP-1 modification |
| CN110004180A (en) * | 2019-04-16 | 2019-07-12 | 上海海洋大学 | A kind of genophore and its preparation method and application modified by Functional Polypeptides A25 |
| CN111154002A (en) * | 2020-01-10 | 2020-05-15 | 上海海洋大学 | Bifunctional peptide K14, gene vector and co-drug delivery system |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102391509A (en) * | 2011-11-29 | 2012-03-28 | 中国人民解放军第二军医大学 | Polyethylenimine derivative, preparation method thereof and application thereof |
| CN102399267A (en) * | 2011-11-22 | 2012-04-04 | 上海海洋大学 | Gene vector modified by bifunctional peptide and preparation method thereof |
| CN103665169A (en) * | 2013-11-21 | 2014-03-26 | 上海海洋大学 | Three-function peptide-modified gene carrier as well as preparation method and application thereof |
| CN104530188A (en) * | 2014-12-11 | 2015-04-22 | 上海海洋大学 | Gene vector modified by functional peptide R11 as well as preparation method and application of gene vector |
-
2015
- 2015-05-15 CN CN201510248910.6A patent/CN104830903B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102399267A (en) * | 2011-11-22 | 2012-04-04 | 上海海洋大学 | Gene vector modified by bifunctional peptide and preparation method thereof |
| CN102391509A (en) * | 2011-11-29 | 2012-03-28 | 中国人民解放军第二军医大学 | Polyethylenimine derivative, preparation method thereof and application thereof |
| CN103665169A (en) * | 2013-11-21 | 2014-03-26 | 上海海洋大学 | Three-function peptide-modified gene carrier as well as preparation method and application thereof |
| CN104530188A (en) * | 2014-12-11 | 2015-04-22 | 上海海洋大学 | Gene vector modified by functional peptide R11 as well as preparation method and application of gene vector |
Non-Patent Citations (1)
| Title |
|---|
| L ROTH ET AL.: "transtumoral targeting enabled by a novel neuropilin-binding peptide", 《ONCOGENE》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109310729A (en) * | 2016-03-31 | 2019-02-05 | 慕尼黑大学 | For improving the synthesis compound of transfection efficiency |
| CN109310729B (en) * | 2016-03-31 | 2022-06-28 | 慕尼黑大学 | Synthetic compounds for improved transfection efficiency |
| CN106480092A (en) * | 2016-10-20 | 2017-03-08 | 上海海洋大学 | A kind of PEGization Fe being modified based on Functional Polypeptides R93+/ PEI genophore and its preparation method and application |
| CN106480092B (en) * | 2016-10-20 | 2019-08-27 | 上海海洋大学 | A kind of PEGylated Fe based on Functional Polypeptides R9 modification3+/ PEI genophore and its preparation method and application |
| CN106801068A (en) * | 2017-01-12 | 2017-06-06 | 西安交通大学 | A kind of non-viral gene vector of autofluorescence degradable poly citrate and preparation method thereof |
| CN109486827A (en) * | 2018-12-04 | 2019-03-19 | 南京林业大学 | A kind of apoferritin nanocages and preparation method thereof of tumor-homing cell-penetrating peptide tLyP-1 modification |
| CN110004180A (en) * | 2019-04-16 | 2019-07-12 | 上海海洋大学 | A kind of genophore and its preparation method and application modified by Functional Polypeptides A25 |
| CN110004180B (en) * | 2019-04-16 | 2023-08-18 | 上海海洋大学 | Gene vector modified by functional peptide A25 and preparation method and application thereof |
| CN111154002A (en) * | 2020-01-10 | 2020-05-15 | 上海海洋大学 | Bifunctional peptide K14, gene vector and co-drug delivery system |
| CN111154002B (en) * | 2020-01-10 | 2022-11-22 | 上海海洋大学 | Bifunctional peptide K14, gene vector and co-drug delivery system |
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