29 research outputs found
Scientific and medical basis of CRISPR/CAS9 and genome editing
No full textDie Verwendung von Stammzellen, iPS-Zellen sowie der Einsatz gentechnischer Methoden wie dem Genome Editing stellen für die Regenerative Medizin prinzipiell Möglichkeiten für die Entwicklung neuer Therapien dar. Die Umsetzung solcher Therapien wirft nicht nur naturwissenschaftlich-technische und medizinische Fragen auf, sondern auch ethische, rechtliche, soziale und ökonomische. Die erfolgreiche Realisierung regenerativmedizinischer Therapien ist daher auf eine disziplinübergreifende Herangehensweise angewiesen. Im Rahmen eines interdisziplinären und internationalen Symposiums sowie einer BMBF-Klausurwoche wurden die Aspekte der Verwendung von Stammzellen, iPS-Zellen sowie des Genome Editings diskutiert. Im Mittelpunkt standen u.a. die arzneimittelrechtliche Handhabung, verfassungsrechtliche und philosophische Fragen in Bezug auf die Kommerzialisierbarkeit menschlicher Körpersubstanzen sowie patentrechtliche Fragen im Umgang mit menschlichen Stammzellen und der Verfahren des Genome Editings.
Mit Beiträgen von
Insa S. Schröder; Susanne Müller und Timo Faltus; Inesa Chmurec; Tereza Hendl; Calvin Wai Loon Ho; Kalina Kamenova; Delphine Pichereau und Emmanuelle Rial-Sebbag; Hannah Schickl; Elena Buglo und Stephan Zuchner; Jochen Taupitz und Juliane Boscheinen; Winfried Kluth; Susanne Beck und Frederike Seitz; Timo Faltus; Rosario Isasi; Katrin Vohland, Julia Diekämper, Alexandra Moormann, Tobias Nettke und Wiebke Rössig; Hans Zillmann und Matthias Kaufmann; Ulrich Storz; Timo Faltus
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Establishing CRISPR Models of Rare Hereditary Neurodegenerative Diseases
No full textRare hereditary neurodegenerative diseases are a group of genetic Mendelian disorders afflicting the central and/or peripheral nervous system. On one hand, these diseases are highly heterogeneous, being derived from a multitude of biochemical pathways and molecular mechanisms. On the other hand, the clinical manifestations may differ significantly between the affected individuals with the same genetic diagnosis. The central theme of this dissertation is neurodegenerative diseases that affect motor systems, such as Charcot-Marie-Tooth (CMT) disease, Hereditary Spastic Paraplegia (HSP) and cerebellar ataxias (CATX). The understanding of genetic disease etiology and its pathogenesis has been precipitated by the discovery of causative disease genes, that were validated in functional models. To date, approximately 60 percent of rare motor system disease causing genetic variants remain unknown, and further genetic discoveries are anticipated to clarify the genetic diagnosis and give rise to gene therapies. Thus, there is an urgent need in the clinical, neurogenetics, and neuroscience fields to quickly identify relevant genes from a large list of candidates and to functionally access them in order to move forward towards therapeutic discoveries. This dissertation aims to elucidate an efficient approach to functional genetic modeling of the human hereditary degenerative motor diseases in zebrafish. Here we show an approach for a time- and cost-effective generation of zebrafish mutants using a novel genetic editing CRISPR technology, followed by a rapid analysis of the relevant motoneuron phenotypes. Furthermore, we describe challenges associated with creating stable animal knockout models, such as complications introduced by the phenomenon of genetic compensation. We highlight that our functional modeling approach opens the doors to a prompt development of targeted gene therapies and gives hope for elimination or partial alleviation of the neurodegenerative processes. Thus, a rapid and efficient gene discovery paradigm presented by current work will have a meaningful impact on the prognosis of disease and improving lives of patients.</div
Erratum: A CADM3 variant causes Charcot-Marie-Tooth disease with marked upper limb involvement (Brain (2021) 144:4 (1197-1213) DOI: 10.1093/brain/awab019)
No full textThe authors and publishers apologize for errors in the author affiliations and the References section. These have been corrected
Biallelic expansion of an intronic repeat in RFC1 is a common cause of late-onset ataxia (vol 51, pg 649, 2019)
No full textIn the version of this article initially published, the name of author Wai Yan Yau was misspelled. The error has been corrected in the HTML and PDF versions of the article
Function Over Form: Modeling Groups of Inherited Neurological Conditions in Zebrafish
No full textZebrafish are a unique cell to behavior model for studying the basic biology of human inherited neurological conditions. Conserved vertebrate genetics and optical transparency provide
in vivo
access to the developing nervous system as well as high-throughput approaches for drug screens. Here we review zebrafish modeling for two broad groups of inherited conditions that each share genetic and molecular pathways and overlap phenotypically: neurodevelopmental disorders such as Autism Spectrum Disorders (ASD), Intellectual Disability (ID) and Schizophrenia (SCZ), and neurodegenerative diseases, such as Cerebellar Ataxia (CATX), Hereditary Spastic Paraplegia (HSP) and Charcot-Marie Tooth Disease (CMT). We also conduct a small meta-analysis of zebrafish orthologs of high confidence neurodevelopmental disorder and neurodegenerative disease genes by looking at duplication rates and relative protein sizes. In the past zebrafish genetic models of these neurodevelopmental disorders and neurodegenerative diseases have provided insight into cellular, circuit and behavioral level mechanisms contributing to these conditions. Moving forward, advances in genetic manipulation, live imaging of neuronal activity and automated high-throughput molecular screening promise to help delineate the mechanistic relationships between different types of neurological conditions and accelerate discovery of therapeutic strategies
Autoimmune Reaction Associated With Long COVID Syndrome and Cardiovascular Disease: A Genetic Case Report
No full textA 35-year-old woman with history of cardiovascular disease presented with shortness of breath, lightheadedness, fatigue, chest pain, and premature ventricular contractions 3 weeks after her second COVID-19 vaccine. Symptoms subsided following catheter ablation and ibuprofen except for chest pain and fatigue, which persisted following ablation and subsequent SARS-CoV-2 infection. The case suggests causal associations between COVID-19 vaccine/infection and recurrence of cardiovascular disease, including long-COVID–like symptoms. (Level of Difficulty: Advanced.
Erratum: Truncating Mutations in UBAP1 Cause Hereditary Spastic Paraplegia (The American Journal of Human Genetics (2019) 104(4) (767–773), (S0002929719300977), (10.1016/j.ajhg.2019.03.001))
No full text(The American Journal of Human Genetics 104, 767–773; April 4, 2019) In the originally published version of this article, authors Ivailo Tournev and Teodora Chamova were mistakenly omitted from the author list. Their names have been added here. The online version of the full article now appears correctly and includes affiliations for the added authors as well as corrections to some of the other affiliations. The authors regret these omissions
Genetic compensation in a stable <i>slc25a46</i> mutant zebrafish: A case for using F0 CRISPR mutagenesis to study phenotypes caused by inherited disease
No full textA phenomenon of genetic compensation is commonly observed when an organism with a disease-bearing mutation shows incomplete penetrance of the disease phenotype. Such incomplete phenotypic penetrance, or genetic compensation, is more commonly found in stable knockout models, rather than transient knockdown models. As such, these incidents present a challenge for the disease modeling field, although a deeper understanding of genetic compensation may also hold the key for novel therapeutic interventions. In our study we created a knockout model of slc25a46 gene, which is a recently discovered important player in mitochondrial dynamics, and deleterious mutations in which are known to cause peripheral neuropathy, optic atrophy and cerebellar ataxia. We report a case of genetic compensation in a stable slc25a46 homozygous zebrafish mutant (hereafter referred as “mutant”), in contrast to a penetrant disease phenotype in the first generation (F0) slc25a46 mosaic mutant (hereafter referred as “crispant”), generated with CRISPR/Cas-9 technology. We show that the crispant phenotype is specific and rescuable. By performing mRNA sequencing, we define significant changes in slc25a46 mutant’s gene expression profile, which are largely absent in crispants. We find that among the most significantly altered mRNAs, anxa6 gene stands out as a functionally relevant player in mitochondrial dynamics. We also find that our genetic compensation case does not arise from mechanisms driven by mutant mRNA decay. Our study contributes to the growing evidence of the genetic compensation phenomenon and presents novel insights about Slc25a46 function. Furthermore, our study provides the evidence for the efficiency of F0 CRISPR screens for disease candidate genes, which may be used to advance the field of functional genetics.</div
Zebrafish: A Pharmacogenetic Model for Anesthesia
No full textGeneral anesthetics are small molecules that interact with and effect the function of many different proteins to promote loss of consciousness, amnesia, and sometimes, analgesia. Owing to the complexity of this state transition and the transient nature of these drug/protein interactions, anesthetics can be difficult to study. The zebrafish is an emerging model for the discovery of both new genes required for the response to and side effects of anesthesia. Here we discuss the tools available to manipulate the zebrafish genome, including both genetic screens and genome engineering approaches. Additionally, there are various robust behavior assays available to study anesthetic and other drug responses. These assays are available for single-gene study or high throughput for genetic or drug discovery. Finally, we present a case study of using propofol as an anesthetic in the zebrafish. These techniques and protocols make the zebrafish a powerful model to study anesthetic mechanisms and drug discovery
