A PhD student is sought to investigate the molecular and genetic bases of vocal learning using a range of cutting edge techniques and model systems. The project will ask how this complex behaviour can be encoded at molecular level by investigating genetic mechanisms and genomic factors. The student will receive comprehensive training to use diverse approaches including molecular, cellular and functional assays, design and testing of genetic engineering methods (CRISPR, shRNA etc), viral packaging, transcriptomics, proteomics and in silico genomic approaches. The student will have the opportunity to work with our extraordinary model system – bats. We have been pioneering the study of bats as neurogenetic models and established them to explore the molecular mechanisms underlying vocal learning and to understand the biology and evolution of speech and language. We have recently generated the first successful genetically engineered bats (transient transgenics) and the student will apply the methods developed in the group, as well as develop new transgenic bat models as part of their project. Working with live animals is not a requirement, as the project is predominantly molecular lab based, but there will be the opportunity to work with the animals if it is desired by the student. This model will shed light onto the molecular encoding of mammalian vocal learning and represent a sophisticated model to provide insight into the mechanisms underlying childhood disorders of language. We are a highly interdisciplinary and collaborative lab and the PhD student will work closely with highly supportive lab members and our rich network of interdisciplinary collaborators, many of whom are world leaders in the field. The student will also be encouraged to present their findings at international conferences (in person or online) and may visit the lab(s) of international collaborators for research stays and knowledge exchange. The PI leads an international genomics consortium, www.bat1k, that is a vibrant community of more than 350 members across >50 countries, which provides many opportunities for interaction, training, knowledge exchange and future career opportunities. This project will provide an excellent opportunity for a student with a keen interest in molecular biology to train in both established as well as new cutting-edge methods applicable to most model systems. Training and personal development will be a key aspect of the PhD and we will work with the student to develop a training plan that suits their needs and personal goals. This will include training in scientific methods, but also in personal and professional development (eg. project design and management, communication skills, writing skills, etc) and will be bolstered by the excellent training available from the transferable skills programme at the University of St Andrews. Many of our lab members are also involved in outreach initiatives and we support students to become involved in local, national or international initiatives according to their interests. The project will be hosted in the School of Biology at the University of St Andrews and benefit from interactions across its three internationally renowned research centres; The Scottish Oceans Institute (SOI), Biomedical Sciences Research Complex (BSRC) and Centre for Biological Diversity (CBD). The incredibly rich research environment and excellent facilities present in the School have led to the School of Biology continuing to be scored by the National Student Survey as one of the top biology schools in the UK. In the student satisfaction led survey, The Times and Sunday Times Good University Guide 2022, the University of St Andrews was ranked as the top UK university, evidence of the rich student environment and social and collegiate atmosphere that leads to a highly positive experience for students at St Andrews.

The Institute of Molecular and Clinical Ophthalmology Basel (IOB) is seeking a highly motivated Research Assistant to join the Retinal Organoid Platform. IOB is a research institute combining basic and clinical research. Its mission is to drive innovations in understanding vision and its diseases and develop new therapies for vision loss. It is a place where your expertise will be valued, your abilities challenged, and your knowledge expanded. The Retinal Organoid Platform uses human retinal organoids derived from pluripotent stem cells as models for inherited retinal degeneration. Therefore, the Retinal Organoid Platform is involved in collecting and reprogramming into iPSC cells from patients with retinal disease as a resource for IOB researchers and collaborators around the globe. Furthermore, the Retinal Organoid Platform is introducing precise patient mutations into hiPSC by genome engineering. Your responsibilities: - In vitro culture of human induced pluripotent stem (iPSC) cells and retinal organoids - Gene editing of iPSC by CRISPR/Cas9, and full characterization of mutant cells - Characterization of iPSC and retinal organoids by various histology, molecular biology and microscopy techniques - Vector construction and molecular cloning - Applying and improving new methodologies to enhance the creation of mutant iPSC - Reprogramming of human primary cells to iPSC - Biobanking of primary cells and iPSC - Involvement in lab management and organization - Close collaboration with group members and IOB groups

The field of human biology faces three major technological challenges. Firstly, the causation problem is difficult to address in humans compared to model animals. Secondly, the complexity problem arises due to the lack of a comprehensive cell atlas for the human body, despite its cellular composition. Lastly, the heterogeneity problem arises from significant variations in both genetic and environmental factors among individuals. To tackle these challenges, we have developed innovative approaches. These include 1) mammalian next-generation genetics, such as Triple CRISPR for knockout (KO) mice and ES mice for knock-in (KI) mice, which enables causation studies without traditional breeding methods; 2) whole-body/brain cell profiling techniques, such as CUBIC, to unravel the complexity of cellular composition; and 3) accurate and user-friendly technologies for measuring sleep and awake states, exemplified by ACCEL, to facilitate the monitoring of fundamental brain states in real-world settings and thus address heterogeneity in human.

SCN8A encodes a major voltage-gated sodium channel expressed in CNS and PNS neurons.  Gain-of-function and loss-of-function mutations contribute to  human disorders, most notably Developmental and Epileptic Encephalophy (DEE). More than 600 affected individuals have been reported, with the most common  mechanism of de novo, gain-of-function mutations.  We have developed constitutive  and conditional models of gain- and loss- of function mutations in the mouse and  characterized the effects of on neuronal firing and neurological phenotypes.  Using CRE lines with cellular and developmental specificity, we have probed the effects of activating  mutant alleles in various classes of neurons in the developing and adult mouse.   Most recently, we are testing genetic therapies that reduce the expression  of gain-of-function mutant alleles.  We are comparing the effectiveness of allele specific  oligos (ASOs), viral delivery of shRNAs, and allele-specific targeting of mutant alleles  using Crispr/Cas9 in mouse models of DEE.

The cerebral cortex contains an extraordinary diversity of excitatory projection neuron (PN) and inhibitory interneurons (IN), wired together to form complex circuits. Spatiotemporally coordinated execution of intrinsic molecular programs by PNs and INs and activity-dependent processes, contribute to cortical development and cortical microcircuits formation. Alterations of these delicate processes have often been associated to neurological/neurodevelopmental disorders. However, despite the groundbreaking discovery that spontaneous activity in the embryonic brain can shape regional identities of distinct cortical territories, it is still unclear whether this early activity contributes to define subtype-specific neuronal fate as well as circuit assembly. In this study, we combined in utero genetic perturbations via CRISPR/Cas9 system and pharmacological inhibition of selected ion channels with RNA-sequencing and live imaging technologies to identify the activity-regulated processes controlling the development of different cortical PN classes, their wiring and the acquisition of subtype specific features. Moreover, we generated human induced pluripotent stem cells (iPSCs) form patients affected by a severe, rare and untreatable form of developmental epileptic encephalopathy. By differentiating cortical organoids form patient-derived iPSCs we create human models of early electrical alterations for studying molecular, structural and functional consequences of the genetic mutations during cortical development. Our ultimate goal is to define the activity-conditioned processes that physiologically occur during the development of cortical circuits, to identify novel therapeutical paths to address the pathological consequences of neonatal epilepsies.

Modern day “neurohackers” are radically self-experimenting, attempting genomic modification with CRISPR-Cas9 constructs and electrode insertion into their cortex amongst a host of other things. Institutions wanting to avoid the risks bought on by these procedures, generally avoid involvement with self-experimenting research. Modern day “neurohackers” are radically self-experimenting, attempting genomic modification with CRISPR-Cas9 constructs and electrode insertion into their cortex amongst a host of other things. Institutions wanting to avoid the risks bought on by these procedures, generally avoid involvement with self-experimenting research. But what is the ethical thing to do? Should researchers be allowed or encouraged to self-experiment? Should institutions support or hinder them? Where do you think that this process of self-experimentation could take us? This presentation by Dr Matt Lennon and Professor Zoltan Molnar of the University of Oxford, will explore the history, future and ethics of self-experimentation. It will explore notable examples of self-experimenters including Isaac Newton, Angelo Ruffini and Oliver Sacks and how a number of these pivotal experiments created paradigm shifts in neuroscience. The presentation will open up a forum for all participants to be involved asking key ethical questions around what should and should not be allowed in self-experimentation research.

Glioblastoma cells trigger pharmacoresistant seizures that may promote tumor growth and diminish the quality of remaining life. To define the relationship between growth of glial tumors and their neuronal microenvironment, and to identify genomic biomarkers and mechanisms that may point to better prognosis and treatment of drug resistant epilepsy in brain cancer, we are analyzing a new generation of genetically defined CRISPR/in utero electroporation inborn glioblastoma (GBM) tumor models engineered in mice. The molecular pathophysiology of glioblastoma cells and surrounding neurons and untransformed astrocytes are compared at serial stages of tumor development. Initial studies reveal that epileptiform EEG spiking is a very early and reliable preclinical signature of GBM expansion in these mice, followed by rapidly progressive seizures and death within weeks. FACS-sorted transcriptomic analysis of cortical astrocytes reveals the expansion of a subgroup enriched in pro-synaptogenic genes that may drive hyperexcitability, a novel mechanism of epileptogenesis. Using a prototypical GBM IUE model, we systematically define and correlate the earliest appearance of cortical hyperexcitability with progressive cortical tumor cell invasion, including spontaneous episodes of spreading cortical depolarization, innate inflammation, and xCT upregulation in the peritumoral microenvironment. Blocking this glutamate exporter reduces seizure load. We show that the host genome contributes to seizure risk by generating tumors in a monogenic deletion strain (MapT/tau -/-) that raises cortical seizure threshold. We also show that the tumor variant profile determines epilepsy risk. Our genetic dissection approach sets the stage to broadly explore the developmental biology of personalized tumor/host interactions in mice engineered with novel human tumor mutations in specified glial cell lineages.

Human genes associated with brain-related diseases are being discovered at an accelerating pace. A major challenge is an identification of the mechanisms through which these genes act, and of potential therapeutic strategies. To elucidate such mechanisms in human cells, we established a CRISPR-based platform for genetic screening in human iPSC-derived neurons, astrocytes and microglia. Our approach relies on CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa), in which a catalytically dead version of the bacterial Cas9 protein recruits transcriptional repressors or activators, respectively, to endogenous genes to control their expression, as directed by a small guide RNA (sgRNA). Complex libraries of sgRNAs enable us to conduct genome-wide or focused loss-of-function and gain-of-function screens. Such screens uncover molecular players for phenotypes based on survival, stress resistance, fluorescent phenotypes, high-content imaging and single-cell RNA-Seq. To uncover disease mechanisms and therapeutic targets, we are conducting genetic modifier screens for disease-relevant cellular phenotypes in patient-derived neurons and glia with familial mutations and isogenic controls. In a genome-wide screen, we have uncovered genes that modulate the formation of disease-associated aggregates of tau in neurons with a tauopathy-linked mutation (MAPT V337M). CRISPRi/a can also be used to model and functionally evaluate disease-associated changes in gene expression, such as those caused by eQTLs, haploinsufficiency, or disease states of brain cells. We will discuss an application to Alzheimer’s Disease-associated genes in microglia.