scholarly journals Genetic Code Expansion in Zebrafish Embryos and Its Application to Optical Control of Cell Signaling

2017 ◽  
Vol 139 (27) ◽  
pp. 9100-9103 ◽  
Author(s):  
Jihe Liu ◽  
James Hemphill ◽  
Subhas Samanta ◽  
Michael Tsang ◽  
Alexander Deiters
Keyword(s):  
Cell Signaling ◽  
Genetic Code ◽  
Optical Control ◽  
Zebrafish Embryos ◽  
Genetic Code Expansion

scholarly journals Using a Quadruplet Codon to Expand the Genetic Code of an Animal

2021 ◽  
Author(s):  
Zhiyan Xi ◽  
Lloyd Davis ◽  
Kieran Baxter ◽  
Ailish Tynan ◽  
Angeliki Goutou ◽  
...  
Keyword(s):  
Amino Acids ◽  
Genetic Code ◽  
Optical Control ◽  
Control Of Gene Expression ◽  
Cell Ablation ◽  
Genetic Code Expansion ◽  
Multicellular Organisms ◽  
Direct Incorporation ◽  
Nematode Worm

Genetic code expansion in multicellular organisms is currently limited to the use of repurposed amber stop codons. Here we introduce a system for the use of quadruplet codons to direct incorporation of non-canonical amino acids in vivo in an animal, the nematode worm Caenorhabditis elegans. We develop hybrid pyrrolysyl tRNA variants to incorporate non-canonical amino acids in response to the quadruplet codon UAGA. We demonstrate the efficiency of the quadruplet decoding system by incorporating photocaged amino acids into two proteins widely used as genetic tools. We use photocaged lysine to express photocaged Cre recombinase for the optical control of gene expression and photocaged cysteine to express photo-activatable caspase for light inducible cell ablation. Our approach will facilitate the routine adoption of quadruplet decoding for genetic code expansion in eukaryotic cells and multicellular organisms.

scholarly journals Optical Control of DNA Helicase Function through Genetic Code Expansion

ChemBioChem ◽  
2017 ◽  
Vol 18 (5) ◽  
pp. 466-469 ◽  
Author(s):  
Ji Luo ◽  
Muwen Kong ◽  
Lili Liu ◽  
Subhas Samanta ◽  
Bennett Van Houten ◽  
...  
Keyword(s):  
Genetic Code ◽  
Dna Helicase ◽  
Optical Control ◽  
Genetic Code Expansion

Structural Basis for Genetic-Code Expansion with Various Bulky Lysine Derivatives by an Engineered Pyrrolysyl-tRNA Synthetase

2018 ◽  
Author(s):  
Tatsuo Yanagisawa ◽  
Mitsuo Kuratani ◽  
Eiko Seki ◽  
Nobumasa Hino ◽  
Kensaku Sakamoto ◽  
...  
Keyword(s):  
Genetic Code ◽  
Trna Synthetase ◽  
Structural Basis ◽  
Genetic Code Expansion

scholarly journals Transferability of N-terminal mutations of pyrrolysyl-tRNA synthetase in one species to that in another species on unnatural amino acid incorporation efficiency

Amino Acids ◽  
2020 ◽  
Author(s):  
Thomas L. Williams ◽  
Debra J. Iskandar ◽  
Alexander R. Nödling ◽  
Yurong Tan ◽  
Louis Y. P. Luk ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Genetic Code ◽  
Unnatural Amino Acids ◽  
Trna Synthetase ◽  
Unnatural Amino Acid ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Genetic Code Expansion ◽  
Incorporation Efficiency

AbstractGenetic code expansion is a powerful technique for site-specific incorporation of an unnatural amino acid into a protein of interest. This technique relies on an orthogonal aminoacyl-tRNA synthetase/tRNA pair and has enabled incorporation of over 100 different unnatural amino acids into ribosomally synthesized proteins in cells. Pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNA from Methanosarcina species are arguably the most widely used orthogonal pair. Here, we investigated whether beneficial effect in unnatural amino acid incorporation caused by N-terminal mutations in PylRS of one species is transferable to PylRS of another species. It was shown that conserved mutations on the N-terminal domain of MmPylRS improved the unnatural amino acid incorporation efficiency up to five folds. As MbPylRS shares high sequence identity to MmPylRS, and the two homologs are often used interchangeably, we examined incorporation of five unnatural amino acids by four MbPylRS variants at two temperatures. Our results indicate that the beneficial N-terminal mutations in MmPylRS did not improve unnatural amino acid incorporation efficiency by MbPylRS. Knowledge from this work contributes to our understanding of PylRS homologs which are needed to improve the technique of genetic code expansion in the future.

scholarly journals ASIC Activation Mechanisms Delineated through Genetic Code Expansion

2021 ◽  
Vol 120 (3) ◽  
pp. 338a
Author(s):  
Matthew L. Rook ◽  
Tyler A. Couch ◽  
Jackson Hernandez ◽  
Alison J. Frontier ◽  
David M. MacLean
Keyword(s):  
Genetic Code ◽  
Activation Mechanisms ◽  
Genetic Code Expansion

Context effects of genetic code expansion by stop codon suppression

2018 ◽  
Vol 46 ◽  
pp. 146-155 ◽  
Author(s):  
Yonatan Chemla ◽  
Eden Ozer ◽  
Itay Algov ◽  
Lital Alfonta
Keyword(s):  
Genetic Code ◽  
Context Effects ◽  
Stop Codon ◽  
Genetic Code Expansion
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