ArticleChemistry & biology2011
Directed evolution of a small-molecule-triggered intein with improved splicing properties in mammalian cells.
Article in Chemistry & biology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed, 60 citations in OpenAlex.
- Synthetic Biology Strategies for the Development of Live Attenuated Influenza Viruses: Recent Advances and Applications.Viruses · 2026Review
- Review
- Orthogonal RNA replication enables directed evolution and Darwinian adaptation in mammalian cells.Nature chemical biology · 2025Article
- Nature-inspired protein ligation and its applications.Nature reviews. Chemistry · 2023Review
- Regulated control of virus replication by 4-hydroxytamoxifen-induced splicing.Frontiers in microbiology · 2023Article
- Discovery of potent and versatile CRISPR-Cas9 inhibitors engineered for chemically controllable genome editing.Nucleic acids research · 2022Article
- Inteins as Drug Targets and Therapeutic Tools.Frontiers in molecular biosciences · 2022Review
- A systematic approach to inserting split inteins for Boolean logic gate engineering and basal activity reduction.Nature communications · 2021Article
- The Effects of Self-cleavage Intein-ELK16 Tag in the Transcript Steric Hindrance of IFN.Indian journal of clinical biochemistry : IJCB · 2021Article
- Directed evolution in mammalian cells.Nature methods · 2021Review
- Construction and Quantitation of a Selectable Protein Splicing Sensor Using Gibson Assembly and Spot Titers.Current protocols · 2021Article
- Inteins in Science: Evolution to Application.Microorganisms · 2020Review
- The crystal structure of the naturally split gp41-1 intein guides the engineering of orthogonal split inteins from cis-splicing inteins.The FEBS journal · 2020Article
- Proximity Induced Splicing Utilizing Caged Split Inteins.Journal of the American Chemical Society · 2019Article
- Off-Pathway-Sensitive Protein-Splicing Screening Based on a Toxin/Antitoxin System.Chembiochem : a European journal of chemical biology · 2019Article
- CRISPR directed evolution of the spliceosome for resistance to splicing inhibitors.Genome biology · 2019Article
- Drug Inducible CRISPR/Cas Systems.Computational and structural biotechnology journal · 2019Review
- An Adaptable Platform for Directed Evolution in Human Cells.Journal of the American Chemical Society · 2018Article
- Protein Engineering Strategies to Expand CRISPR-Cas9 Applications.International journal of genomics · 2018Review
- Intein Zymogens: Conditional Assembly and Splicing of Split Inteins via Targeted Proteolysis.Journal of the American Chemical Society · 2017Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Laboratory-created small-molecule-dependent inteins enable protein structure and function to be controlled posttranslationally in living cells. Previously we evolved inteins that splice efficiently in Saccharomyces cerevisiae only in the presence of the cell-permeable small molecule 4-hydroxytamoxifen (4-HT). In mammalian cells, however, these inteins exhibited lower splicing efficiencies and slower splicing in the presence of 4-HT, as well as higher background splicing in the absence of 4-HT. Here we further evolved ligand-dependent inteins in yeast at 30°C and 37°C. The resulting second-generation evolved inteins exhibit substantially improved splicing yields and kinetics. The improvements carried over to mammalian cells, in which the newly evolved inteins spliced with substantially greater (∼2- to 8-fold) efficiency while maintaining low background splicing levels. These second-generation inteins augment the promise of ligand-dependent protein splicing for probing protein function in mammalian cells.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.