Xenopus Oocyte Expression Assay

RNA was transcribed using the SP6 mMessageMachine kit (Thermofisher, Milan, Italy) after linearization with MluI. Xenopus laevis oocytes were injected with ~6 ng of RNA and incubated at 18 °C for 2–5 days prior to measurements as described previously [26 (link)].

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Palmer E.E., Pusch M., Picollo A., Forwood C., Nguyen M.H., Suckow V., Gibbons J., Hoff A., Sigfrid L., Megarbane A., Nizon M., Cogné B., Beneteau C., Alkuraya F.S., Chedrawi A., Hashem M.O., Stamberger H., Weckhuysen S., Vanlander A., Ceulemans B., Rajagopalan S., Nunn K., Arpin S., Raynaud M., Motter C.S., Ward-Melver C., Janssens K., Meuwissen M., Beysen D., Dikow N., Grimmel M., Haack T.B., Clement E., McTague A., Hunt D., Townshend S., Ward M., Richards L.J., Simons C., Costain G., Dupuis L., Mendoza-Londono R., Dudding-Byth T., Boyle J., Saunders C., Fleming E., El Chehadeh S., Spitz M.A., Piton A., Gerard B., Abi Warde M.T., Rea G., McKenna C., Douzgou S., Banka S., Akman C., Bain J.M., Sands T.T., Wilson G.N., Silvertooth E.J., Miller L., Lederer D., Sachdev R., Macintosh R., Monestier O., Karadurmus D., Collins F., Carter M., Rohena L., Willemsen M.H., Ockeloen C.W., Pfundt R., Kroft S.D., Field M., Laranjeira F.E., Fortuna A.M., Soares A.R., Michaud V., Naudion S., Golla S., Weaver D.D., Bird L.M., Friedman J., Clowes V., Joss S., Pölsler L., Campeau P.M., Blazo M., Bijlsma E.K., Rosenfeld J.A., Beetz C., Powis Z., McWalter K., Brandt T., Torti E., Mathot M., Mohammad S.S., Armstrong R, & Kalscheuer V.M. (2022). Functional and clinical studies reveal pathophysiological complexity of CLCN4-related neurodevelopmental condition. Molecular Psychiatry, 28(2), 668-697.

Publication 2022

SP6 mMessageMachine kit

Oocytes Xenopus laevis

Corresponding Organization : Instituto Biofisika

Other organizations : UNSW Sydney, Max Planck Institute for Molecular Genetics, Lebanese American University, Génétique Médicale & Génomique Fonctionelle, Centre Hospitalier Universitaire de Nantes, Nantes Université, King Faisal Specialist Hospital & Research Centre, VIB-UAntwerp Center for Molecular Neurology, Ghent University Hospital, Antwerp University Hospital, University of Antwerp, Liverpool Hospital, Children's Hospital at Westmead, Sydney Children’s Hospitals Network, Centre Hospitalier Universitaire de Tours, Akron Children's Hospital, Heidelberg University, University of Tübingen, Great Ormond Street Hospital, University College London, Princess Anne Hospital, King Edward Memorial Hospital, Washington University in St. Louis, Murdoch Children's Research Institute, Hospital for Sick Children, Hunter Genetics, Children's Mercy Hospital, Hôpitaux Universitaires de Strasbourg, Hôpital Civil, Strasbourg, Haukeland University Hospital, University of Manchester, Columbia University Irving Medical Center, Texas Tech University Health Sciences Center, Integrative Health Technologies (United States), Hillcrest Clinics, Institute of Pathology and Genetics, Royal Prince Alfred Hospital, Children's Hospital of Eastern Ontario, San Antonio Military Medical Center, Radboud University Nijmegen, Radboud University Medical Center, Pluryn, Centro Hospitalar do Porto, Universidade do Porto, Centre Hospitalier Universitaire de Bordeaux, Indiana University – Purdue University Indianapolis, Rady Children's Hospital-San Diego, University of California, San Diego, London North West Healthcare NHS Trust, Queen Elizabeth University Hospital, Universitair Ziekenhuis Brussel, Vrije Universiteit Brussel, Centre Hospitalier Universitaire Sainte-Justine, Université de Montréal, Texas A&M Health Science Center, Texas A&M University, Leiden University, Baylor College of Medicine, Centogene (Germany), Ambry Genetics (United States), CHU Dinant Godinne UCL Namur, Addenbrooke's Hospital

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Variable analysis

independent variables

Linearization of RNA with MluI

dependent variables

Xenopus laevis oocyte response to injected RNA

control variables

Amount of injected RNA (~6 ng)
Incubation temperature (18 °C)
Incubation duration (2-5 days)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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