ArticleProceedings of the National Academy of Sciences of the United States of America2017
Structure-guided SCHEMA recombination generates diverse chimeric channelrhodopsins.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 62 citations in OpenAlex.
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- Optimizing enzyme thermostability by combining multiple mutations using protein language model.mLife · 2024Article
- Rhodopsin-based light-harvesting system for sustainable synthetic biology.Microbial biotechnology · 2024Article
- Opportunities and Challenges for Machine Learning-Assisted Enzyme Engineering.ACS central science · 2024Review
- Selection pressures on evolution of ribonuclease H explored with rigorous free-energy-based design.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Computational Protein Design - Where it goes?Current medicinal chemistry · 2024Review
- Channelrhodopsin fluorescent tag replacement for clinical translation of optogenetic hearing restoration.Molecular therapy. Methods & clinical development · 2023Article
- A kinetic-optimized CoChR variant with enhanced high-frequency spiking fidelity.Biophysical journal · 2022Article
- Engineering rhodopsins' activation spectra using a FRET-based approach.Biophysical journal · 2022Article
- Rhodopsins: An Excitingly Versatile Protein Species for Research, Development and Creative Engineering.Frontiers in chemistry · 2022Review
- Facilitating Machine Learning-Guided Protein Engineering with Smart Library Design and Massively Parallel Assays.Advanced genetics (Hoboken, N.J.) · 2021Article
- Improved RAD51 binders through motif shuffling based on the modularity of BRC repeats.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Development of an optogenetic gene sensitive to daylight and its implications in vision restoration.NPJ Regenerative medicine · 2021Article
- Artificial Intelligence, Machine Learning and Deep Learning in Ion Channel Bioinformatics.Membranes · 2021Review
- "NAD-display": Ultrahigh-Throughput in Vitro Screening of NAD(H) Dehydrogenases Using Bead Display and Flow Cytometry.Angewandte Chemie (International ed. in English) · 2021Article
- Emerging Diversity of Channelrhodopsins and Their Structure-Function Relationships.Frontiers in cellular neuroscience · 2021Review
- Split & mix assembly of DNA libraries for ultrahigh throughput on-bead screening of functional proteins.Nucleic acids research · 2020Article
- Synthetic chimeric nucleases function for efficient genome editing.Nature communications · 2019Article
- Machine learning-guided channelrhodopsin engineering enables minimally invasive optogenetics.Nature methods · 2019Article
- Microbial Experimental Evolution - a proving ground for evolutionary theory and a tool for discovery.EMBO reports · 2019Review
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
Abstract
Integral membrane proteins (MPs) are key engineering targets due to their critical roles in regulating cell function. In engineering MPs, it can be extremely challenging to retain membrane localization capability while changing other desired properties. We have used structure-guided SCHEMA recombination to create a large set of functionally diverse chimeras from three sequence-diverse channelrhodopsins (ChRs). We chose 218 ChR chimeras from two SCHEMA libraries and assayed them for expression and plasma membrane localization in human embryonic kidney cells. The majority of the chimeras express, with 89% of the tested chimeras outperforming the lowest-expressing parent; 12% of the tested chimeras express at even higher levels than any of the parents. A significant fraction (23%) also localize to the membrane better than the lowest-performing parent ChR. Most (93%) of these well-localizing chimeras are also functional light-gated channels. Many chimeras have stronger light-activated inward currents than the three parents, and some have unique off-kinetics and spectral properties relative to the parents. An effective method for generating protein sequence and functional diversity, SCHEMA recombination can be used to gain insights into sequence-function relationships in MPs.
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Registered trials
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.