How to use stem ring: Analysis of hot topics and hot content on the entire network in the past 10 days
Recently, the popularity of stem-loop structure in the fields of molecular biology and genetic engineering has continued to rise, especially in the application of CRISPR technology and RNA vaccine research and development has caused widespread discussion. This article will combine popular topics across the network for the past 10 days to analyze the uses, technical points and latest research progress of stem rings.
1. Basic concepts of stem and ring structure
Stem rings are secondary structures formed by single-stranded nucleic acid molecules (DNA or RNA) through base pairing, includingStem(Double-linked area) andThe ring part(Unpaired single-stranded area). Its functions are diverse, involving gene expression regulation, RNA stability maintenance, etc.
| Structural parts | Functional Features | Application scenarios |
|---|---|---|
| Stem | Base complementary pairing provides structural stability | CRISPR guided RNA design |
| The ring part | Flexible area, involved in molecular recognition | miRNA targeted binding |
2. Popular application cases in the past 10 days
1.RNA vaccine optimization(Popularity ★★★★★☆)
Moderna's latest research shows that by adjusting the structure of the 5' end stem ring of mRNA vaccine, protein expression efficiency can be improved by 40%.
2.Gene editing tool upgrade(Popularity ★★★★★★★)
The "CRISPR-SL" system developed by the Broad Research Institute uses stem ring structure to achieve a breakthrough in editing accuracy:
| Technical Indicators | Traditional CRISPR | Stem ring optimization version |
|---|---|---|
| Off-target rate | 15-25% | ≤3% |
| Editing efficiency | 70-80% | 92-95% |
3. Experimental operation guide
Design points:
• Stem length: 4-8bp recommended
• Ring size: usually 4-10nt
• GC content: controlled at 40-60%
Comparison of common tools:
| Software name | Advantages | Applicable scenarios |
|---|---|---|
| RNAfold | Free online tools | Quickly predict secondary structure |
| NUPACK | Multimolecular interaction analysis | Complex system design |
4. Industry frontier trends
1. Shanghai Jiaotong University team discovered new typesPhotoresponsive stem ring switch, can realize the time and air conditioning of gene expression (Nature Biotechnology, 2023)
2. The latest guide of the US FDA emphasizes that the stem ring structure of RNA drugs needs to be used asKey Quality Attributes(CQA) for characterization
5. Frequently Asked Questions
Q: Will stem ring structure affect PCR amplification efficiency?
A: Yes. It is recommended to avoid the primer binding region or use hot-start Taq enzyme.
Q: How to verify the designed stem ring structure?
A: Recommended three-step method: ① Software prediction ② Electrophoretic mobility experiment (EMSA) ③ Enzyme protection experiment
Conclusion
With the development of synthetic biology, stem ring structure has become a core tool in the design of gene components. Mastering its design principles and the latest technical trends will help scientific researchers make breakthroughs in gene therapy, synthetic biology and other fields.
check the details
check the details
en
