Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Application of long chain non-coded RNA-NKILA in bone tissue damage repair

A long-chain non-coding, damage repair technology, applied in the field of stem cell bioengineering, can solve problems such as many complications, immune rejection, iatrogenic infection, etc., and achieve the effect of promoting calcium ion deposition and enhancing alkaline phosphatase activity

Active Publication Date: 2019-01-04
XINXIANG MEDICAL UNIV
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For sports-induced bone injuries, traditional methods such as autologous bone grafting, allogeneic bone grafting, blood-supplied free bone grafting, and biomaterial replacement are often used in the medical field. Rejection and iatrogenic infection and many other problems, the treatment effect is not ideal

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of long chain non-coded RNA-NKILA in bone tissue damage repair
  • Application of long chain non-coded RNA-NKILA in bone tissue damage repair
  • Application of long chain non-coded RNA-NKILA in bone tissue damage repair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Acquisition of NKILA gene fragments

[0030] According to the RNA nucleotide sequence of the long-chain non-coding RNA-NKILA in the NCBI database (RefSeq sequence number: NR_131157.1, as shown in SEQ ID NO: 1 in the sequence listing) and BamH I and Xba I on the lentiviral vector pLV Specific enzyme cutting sites, using DNA chem192 synthesizer, according to the principle of solid-phase phosphoramidite triester method, double-strand synthesis of NKILA gene fragments, and adding two specific enzyme cutting sites on the 5' end and 3' end respectively site BamH I (GGATCC) and Xba I (TCTAGA) to obtain a NKILA gene fragment containing a specific restriction site, the nucleotide sequence of which is shown in SEQ ID NO: 2 in the sequence listing.

Embodiment 2

[0031] Example 2: Construction of lentiviral expression vector pLV-NKILA

[0032] Restriction endonucleases BamH I and Xba I were used to double-enzyme the lentiviral empty vector pLV (its nucleotide sequence is shown in SEQ ID NO: 3 in the sequence listing) and the NKILA gene fragment obtained in Example 1, respectively. The T4 DNA ligase system was used to carry out ligation reaction between the digested NKILA gene fragment and the linear lentiviral empty vector pLV, then transform competent cells, screen positive colonies, and extract the plasmids of positive colonies to obtain the lentiviral expression vector pLV- NKILA.

[0033] 1. Enzyme digestion of lentiviral empty vector pLV

[0034] The enzyme digestion reaction system is as follows (20 μL):

[0035]

[0036] Reaction conditions for enzyme cleavage: react at 37°C for 4 hours.

[0037] 2. Enzyme digestion of NKILA gene fragment

[0038] The enzyme digestion reaction system is as follows (20 μL):

[0039]

...

Embodiment 3

[0054] Example 3: Lentiviral Packaging

[0055] HEK293T cells were seeded in six-well culture plates for culture, and cultured in a cell culture incubator at 37°C with DMEM complete medium containing 10% fetal bovine serum (FBS). When the cell density reached about 70%, the Lipofectamine 3000 liposome transfection reagent (purchased from Thermo Fisher, model L3000015), respectively transfected lentiviral expression vector pLV-NKILA and lentiviral empty vector pLV into HEK293T cells for lentiviral packaging. Specific steps are as follows:

[0056] 1) Take two 1.5mL EP tubes and add 150μL serum-free DMEM medium to each;

[0057] 2) Add 1 μg lentiviral expression vector pLV-NKILA, 0.5 μg lentiviral packaging plasmid pSPAX2, and 0.5 μg lentiviral packaging plasmid pMD2G into one of the EP tubes, and mix well;

[0058] 3) Add Lipofectamine 3000 liposome transfection reagent into another EP tube and mix well;

[0059] 4) Quickly add the plasmid solution obtained in step 2) to the...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides application of long chain non-coded RNA-NKILA in bone tissue damage repair. Experiments verify that overexpressed NKILA can promote calcium ion deposition in a mesenchymal stemcell obviously, enhance the activity of alkaline phosphatase in the mesenchymal stem cell obviously and improve expression quantities of osteogenic differentiation markers RUNX2, SP7 and SPP1 genes inthe mesenchymal stem cell obviously.

Description

technical field [0001] The invention belongs to the technical field of stem cell bioengineering, and more specifically relates to the application of long-chain non-coding RNA-NKILA in repairing bone tissue damage. Background technique [0002] Bone injury repair and regeneration has always been a major problem in the medical field. For sports-induced bone injuries, traditional methods such as autologous bone grafting, allogeneic bone grafting, blood-supplied free bone grafting, and biomaterial replacement are often used in the medical field. Rejection and iatrogenic infection and many other problems, the treatment effect is not satisfactory. With the continuous development of bone tissue engineering technology, people gradually began to use tissue engineering methods to solve the problem of bone tissue damage repair. As a kind of seed cells with good value-proliferating ability and multi-directional differentiation potential, mesenchymal stem cells have come into people's ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/867C12N5/10C12Q1/6883
CPCC12N15/113C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 林俊堂朱鑫星于金金钟根深杨芬冯岩岩
Owner XINXIANG MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com