Synthetic Biology seminars

Natural proteins solve problems shaped by evolution. David Baker describes a different approach: designing proteins from scratch to meet current challenges in health, technology and sustainability. His laboratory develops deep-learning methods that generate proteins with new functions, synthesizes genes encoding the designs, and experimentally characterizes the resulting molecules. The lecture explains how this design-build-test approach extends beyond modifying existing natural proteins and presents applications of computational protein design to contemporary problems. Online via the Royal Society public livestream. The lecture is free to watch live on the Royal Society’s YouTube channel.

Jennifer Brophy presents two methods for programming when and where genes are expressed in plants and their associated microbes. One creates a chemical signal using the noncanonical amino acid O-methyl-L-tyrosine. Engineered Arabidopsis, tomato and poplar produce the signal, while engineered Bacillus subtilis and soil bacteria respond by activating gene expression. Restricting signal production to selected plant tissues allows local control of microbial activity. The second approach builds a genetic circuit that counts successive branching events in roots. It produces different expression patterns at different branch orders, helping researchers distinguish the contributions of roots that otherwise have similar molecular identities. Together, the approaches support experiments on plant development and the engineering of plant–microbe partnerships.

UC Berkeley Molecular & Cell Biology seminar (Divisions of Biochemistry, Biophysics & Structural Biology; Cell Biology, Development & Physiology; and Genetics, Genomics, Evolution & Development) by Luke Gilbert of UCSF on repurposing CRISPR systems to activate and silence genes.

David Baker presents recent advances in designing proteins for functions that natural evolution has not been required to solve. The approach begins with a desired structure or activity and uses deep learning to propose amino acid sequences expected to realise it. The resulting designs are translated into synthetic genes, produced as proteins and tested experimentally. The lecture connects computational design with laboratory characterisation, explaining how the combination can create new molecular functions relevant to medicine, technology and sustainability. Its focus is the construction and evaluation of proteins from scratch, extending the protein-design research recognised by the 2024 chemistry prize.

This lecture examines the sequence features through which enhancers bind transcription factors and control gene expression. It introduces computational approaches, including neural networks, for recognising regulatory DNA and potentially designing synthetic elements. It also compares recurring arrangements of enhancers around genes important in mammalian development.

A key question for basic biology and regenerative medicine concerns the way in which evolution exploits physics toward adaptive form and function. While genomes specify the molecular hardware of cells, what algorithms enable cellular collectives to reliably build specific, complex, target morphologies? Our lab studies the way in which all cells, not just neurons, communicate as electrical networks that enable scaling of single-cell properties into collective intelligences that solve problems in anatomical feature space. By learning to read, interpret, and write bioelectrical information in vivo, we have identified some novel controls of growth and form that enable incredible plasticity and robustness in anatomical homeostasis. In this talk, I will describe the fundamental knowledge gaps with respect to anatomical plasticity and pattern control beyond emergence, and discuss our efforts to understand large-scale morphological control circuits. I will show examples in embryogenesis, regeneration, cancer, and synthetic living machines. I will also discuss the implications of this work for not only regenerative medicine, but also for fundamental understanding of the origin of bodyplans and the relationship between genomes and functional anatomy.

Developmental and epileptic encephalopathies (DEEs) are a broad spectrum of genetic epilepsies associated with impaired neurological development as a direct consequence of a genetic mutation, in addition to the effect of the frequent epileptic activity on brain. Compelling genetic studies indicate that heterozygous de novo mutations represent the most common underlying genetic mechanism, in accordance with the sporadic presentation of DEE. De novo mutations may exert a loss-of-function (LOF) on the protein by decrementing expression level and/or activity, leading to functional haploinsufficiency. These diseases share several features: severe and frequent refractory seizures, diffusely abnormal background activity on EEG, intellectual disability often profound, and severe consequences on global development. One of major causes of early onset DEE are de novo heterozygous mutations in syntaxin-binding-protein-1 gene STXBP1, which encodes a membrane trafficking protein playing critical role in vesicular docking and fusion. LOF STXBP1 mutations lead to a failure of neurotransmitter secretion from synaptic vesicles. Core clinical features of STXBP1 encephalopathy include early-onset epilepsy with hypsarrhythmic EEG, or burst-suppression pattern, or multifocal epileptiform activity. Seizures are often resistant to standard treatments and patients typically show intellectual disability, mostly severe to profound. Additional neurologic features may include autistic traits, movement disorders (dyskinesia, dystonia, tremor), axial hypotonia, and ataxia, indicating a broader neurologic impairment. Patients with severe neuro-cognitive features but without epilepsy have been reported. Recently, a new class of natural and synthetic non-coding RNAs have been identified, enabling upregulation of protein translation in a gene-specific way (SINEUPs), without any increase in mRNA of the target gene. SINEUPs are translational activators composed by a Binding Domain (BD) that overlaps, in antisense orientation, to the sense protein-coding mRNA, and determines target selection; and an Effector Domain (ED), that is essential for protein synthesis up regulation. SINEUPs have been shown to restore the physiological expression of a protein in case of haploinsufficiency, without driving excessive overexpression out of the physiological range. This technology brings many advantages, as it mainly acts on endogenous target mRNAs produced in situ by the wild-type allele; this action is limited to mRNA under physiological regulation, therefore no off-site effects can be expected in cells and tissues that do not express the target transcript; by acting only on a posttranscriptional level, SINEUPs do not trigger hereditable genome editing. After bioinformatic analysis of the promoter region of interest, we designed SINEUPs with 3 different BD for STXBP1. Human neurons from iPSCs were treated and STXBP1 levels showed a 1.5-fold increase compared to the Negative control. RNA levels of STXBP1 after the administration of SINEUPs remained stable as expected. These preliminary results proved the SINEUPs potential to specifically increase the protein levels without impacting on the genome. This is an extremely flexible approach to target many developmental and epileptic encephalopathies caused by haploinsufficiency, and therefore to address these diseases in a more tailored and radical way.

Clock networks containing the same central architectures may vary drastically in their potential to oscillate, raising the question of what controls robustness, one of the essential functions of an oscillator. We computationally generate an atlas of oscillators and found that, while core topologies are critical for oscillations, local structures substantially modulate the degree of robustness. Strikingly, two local structures, incoherent and coherent inputs, can modify a core topology to promote and attenuate its robustness, additively. The findings underscore the importance of local modifications to the performance of the whole network. It may explain why auxiliary structures not required for oscillations are evolutionary conserved. We also extend this computational framework to search hidden network motifs for other clock functions, such as tunability that relates to the capabilities of a clock to adjust timing to external cues. Experimentally, we developed an artificial cell system in water-in-oil microemulsions, within which we reconstitute mitotic cell cycles that can perform self-sustained oscillations for 30 to 40 cycles over multiple days. The oscillation profiles, such as period, amplitude, and shape, can be quantitatively varied with the concentrations of clock regulators, energy levels, droplet sizes, and circuit design. Such innate flexibility makes it crucial to studying clock functions of tunability and stochasticity at the single-cell level. Combined with a pressure-driven multi-channel tuning setup and long-term time-lapse fluorescence microscopy, this system enables a high-throughput exploration in multi-dimension continuous parameter space and single-cell analysis of the clock dynamics and functions. We integrate this experimental platform with mathematical modeling to elucidate the topology-function relation of biological clocks. With FRET and optogenetics, we also investigate spatiotemporal cell-cycle dynamics in both homogeneous and heterogeneous microenvironments by reconstructing subcellular compartments.

Gerald Joyce explores attempts to reconstruct features of RNA-based life in the laboratory. Under the RNA world hypothesis, early organisms used RNA to store information and catalyze replication before the emergence of DNA genomes. Directed evolution has produced RNA enzymes that copy diverse templates, synthesize structured and catalytic RNAs, and amplify short templates in an RNA-catalyzed analogue of the polymerase chain reaction. A variant can make DNA from an RNA template, suggesting a possible route between RNA and DNA genomes. Joyce also describes cross-chiral enzymes that synthesize RNA of opposite handedness and can assemble their own enantiomers from fragments. The lecture examines these complementary approaches to replicating functional RNA, their relevance to the origins of life, and potential diagnostic and clinical applications.