Related Experiment Video
Updated: Jul 30, 2025
13:11
Super-Resolution Imaging of Bacterial Secreted Proteins Using Genetic Code Expansion
Published on: February 10, 2023
1.5K
Shedding light on ORAI1 channel with genetic code expansion
Sher Ali1, Guolin Ma2, Yubin Zhou2
1Center for Translational Cancer Research, Institute of Biosciences and Technology, Texas A&M University, Houston, TX, 77030, United States of America.
Cell Calcium
|May 17, 2023
Summary
Researchers used genetic code expansion to insert photocrosslinking amino acids into the ORAI1 calcium channel. This allows for UV light control of calcium influx and downstream signaling in cells.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Genetic code expansion utilizes suppressor tRNA and orthogonal aminoacyl-tRNA synthetase pairs.
- This technology enables the incorporation of unnatural amino acids (UAAs) into proteins.
- UAAs can be used to control protein activity and biological systems.
Purpose of the Study:
- To incorporate photocrosslinking UAAs into the transmembrane domains of ORAI1.
- To enable UV light-inducible calcium influx across the plasma membrane.
- To allow mechanistic interrogation of the calcium release-activated calcium (CRAC) channel and control downstream signaling.
Main Methods:
- Utilized genetic code expansion technology.
- Incorporated photocrosslinking unnatural amino acids into ORAI1 transmembrane domains.
- Applied UV light to induce and control calcium influx.
Main Results:
- Successfully incorporated photocrosslinking UAAs into ORAI1.
- Achieved UV light-inducible calcium influx across the plasma membrane.
- Enabled single amino acid level interrogation of the CRAC channel and control of downstream signaling.
Conclusions:
- Genetic code expansion is a powerful tool for controlling protein function.
- Photocrosslinking UAAs in ORAI1 provide a novel method for studying calcium signaling.
- This approach offers remote control over calcium-modulated pathways in mammalian cells.
Related Concept Videos
02:24Ribosome Profiling
3.6K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.6K
02:32Exon Recombination
3.6K
The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes.
Exon shuffling follows “splice frame rules.” Each exon...
Exon shuffling follows “splice frame rules.” Each exon...
3.6K
02:11Reporter Genes
11.7K
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
11.7K
02:46Genetic Screens
5.0K
Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
5.0K
02:37Gene Duplication and Divergence
6.2K
The seminal work of Ohno in 1970 popularized the idea of gene duplication and divergence. DNA sequence comparison studies reveal that a large portion of the genes in bacteria, archaebacteria, and eukaryotes was generated by gene duplication and divergence, indicating its critical role in evolution.
The duplicated copies of the gene are called Paralogs. Paralogs with similar sequences and functions form a gene family. Across several species, a large number of gene families are...
The duplicated copies of the gene are called Paralogs. Paralogs with similar sequences and functions form a gene family. Across several species, a large number of gene families are...
6.2K
02:28Leaky Scanning
5.2K
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.2K