Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for determining minimum entrapment binding ratio in polypeptide and siRNA co-assembly

A technology of co-assembly and binding ratio, applied in the field of bioengineering, can solve problems such as the minimum encapsulation and binding ratio of peptides and siRNA that are not recorded in experiments.

Pending Publication Date: 2022-06-24
陈璞
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no experimental method has been documented so far on how to determine the minimum loading binding ratio of peptides to siRNA, thus limiting the siRNA approach.

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
  • Method for determining minimum entrapment binding ratio in polypeptide and siRNA co-assembly
  • Method for determining minimum entrapment binding ratio in polypeptide and siRNA co-assembly
  • Method for determining minimum entrapment binding ratio in polypeptide and siRNA co-assembly

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] This embodiment discloses an experimental method for detecting the binding ratio of carrier polypeptide to siRNA, which specifically includes the following steps:

[0060] (1) Prepare 2% agarose gel in advance. The preparation method is as follows: a. Weigh 3.2g of agarose powder into a beaker, add 1×TAE to make the volume to 160mL, shake and mix slightly, and place it in a microwave oven to heat for 4- 7min, until completely dissolved, after cooling for a while, add GelRed Nucleic Acid Gel Stain (BIOTIUM; Cat: 41003) dye to make the working concentration 1×; b. Pour the dissolved agarose into the mold inserted into the comb, and leave it at room temperature for 30min, Pull out the comb; c. Put the prepared gel into the electrophoresis tank, and perform electrophoresis experiment after adding the sample.

[0061] (2) Sample preparation method:

[0062] The CPP and siRNA were configured in a series of ratios as shown in Table 1 below. Polypeptide stock solution concent...

Embodiment 2

[0075] This embodiment discloses an experimental method for detecting the binding ratio of carrier polypeptide to siRNA, which specifically includes the following steps:

[0076] (1) Prepare 2% agarose gel in advance. The preparation method is as follows: a. Weigh 3.2g of agarose powder into a beaker, add 1×TAE to make up to 160ml, shake and mix evenly, and heat in a microwave oven for 4- 7min, until completely dissolved; b. Pour the dissolved agarose into the mold inserted into the comb, leave it at room temperature for 30min, and pull out the comb; c. Put the gel into the electrophoresis tank, and conduct electrophoresis experiment after adding the sample .

[0077] (2) Sample preparation method:

[0078] The CPP and siRNA were configured in a series of ratios as shown in Table 3 below. Polypeptide stock solution concentration: 500 μM; siRNA stock solution concentration: 50 μM solution.

[0079] table 3

[0080]

[0081] The CPP and siRNA were configured in a series o...

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 belongs to the field of bioengineering, and particularly relates to a method for determining the minimum entrapment binding ratio in a polypeptide and siRNA co-assembly. The invention discloses a method for determining the minimum entrapment binding ratio in a polypeptide and siRNA co-assembly. The method comprises the following steps: S1, preparing 2% agarose gel; s2, preparing the CPP-siRNA nano particles; s3, target polypeptide is added, an electrophoresis experiment is carried out, and a gel imager is used for imaging; and observing a map in an imaging result by naked eyes, selecting a group with no tailing of a protein electrophoresis track in the image, and taking the minimum binding ratio in the group without tailing as the minimum entrapment binding ratio. The invention discovers a method for determining the minimum entrapment binding ratio of the positively charged nanoparticles formed by self-assembly of the polypeptide and the siRNA, discovers several polypeptide sequences suitable for the method, and fills the blank in the prior art. The method is simple to operate, and the judgment method is very visual and easy to operate.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a method for determining the minimum encapsulation and binding ratio in the co-assembly of polypeptide and siRNA. [0002] technical background [0003] RNAi (RNA interference) was discovered in the process of studying C. elegans antisense RNA, the degradation process of homologous RNA mediated by dsRNA. RNA interference refers to a phenomenon that is highly conserved during evolution, induced by double-stranded RNA (dsRNA), and homologous mRNA is efficiently and specifically degraded. Gene silencing mainly includes pre-transcriptional gene silencing (TGS) and post-transcriptional gene silencing (PTGS): TGS refers to the abnormal transcription of genes due to DNA modification or chromosomal heterochromatinization; Sequence-specific degradation mechanisms of target mRNAs in the cytoplasm. Sometimes transgenes cause both TGS and PTGS. [0004] Since RNA interference tech...

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): G01N27/447
CPCG01N27/447
Inventor 孙婷婷舒智愚牟泉萌陈璞
Owner 陈璞
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap 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