Stem Cell Research (Feb 2025)
Generation of a genetically encoded voltage indicator MARINA reporter human iPS cell line using Cas9 (VULSCi002-A-2)
- Aistė Petruškevičiūtė,
- Ugnė Šimuliūnaitė,
- Catalina M. Polanco,
- Barbara Rojas,
- Simonas Kuras,
- Beatričė Valatkaitė-Rakštienė,
- Rimvydas Norvilas,
- Akshay Kumar Vijaya,
- Patricia Muñoz,
- Urtė Neniškytė,
- Artūras Jakubauskas,
- Aurelijus Burokas,
- Ivan Nalvarte,
- Jose Inzunza,
- Daniel Naumovas,
- Mindaugas Stoškus,
- Laimonas Griškevičius,
- Daiva Baltriukienė,
- Jonathan Arias
Affiliations
- Aistė Petruškevičiūtė
- VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania; Laboratory of Nuclease Enabled Cell Therapies, Lithuania
- Ugnė Šimuliūnaitė
- VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania; Laboratory of Nuclease Enabled Cell Therapies, Lithuania; Doctoral Program of Biochemistry, Vilnius University Life Science Center, Lithuania
- Catalina M. Polanco
- VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania; Laboratory of Nuclease Enabled Cell Therapies, Lithuania
- Barbara Rojas
- VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania; Laboratory of Nuclease Enabled Cell Therapies, Lithuania
- Simonas Kuras
- VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania; Laboratory of Nuclease Enabled Cell Therapies, Lithuania; Doctoral Program of Biology, Vilnius University Life Science Center, Lithuania
- Beatričė Valatkaitė-Rakštienė
- Hematology, Oncology and Transfusion Medicine Center, Vilnius University Hospital Santaros Klinikos, Vilnius, Lithuania
- Rimvydas Norvilas
- VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania; Laboratory of Nuclease Enabled Cell Therapies, Lithuania; Hematology, Oncology and Transfusion Medicine Center, Vilnius University Hospital Santaros Klinikos, Vilnius, Lithuania
- Akshay Kumar Vijaya
- Doctoral Program of Biochemistry, Vilnius University Life Science Center, Lithuania; Department of Biological Models, Institute of Biochemistry, Life Sciences Center, Vilnius University, Saulėtekio Ave. 7, LT-10257 Vilnius, Lithuania
- Patricia Muñoz
- Karolinska Institutet, Department of Neurobiology, Care Sciences and Society, BioClinicum, Visionsgatan 4, Solna, Sweden; Karolinska Institutet Stem Cell Organoid (KISCO) Facility, Department of Laboratory Medicine, Alfred Nobels allé 8, Huddinge, Sweden
- Urtė Neniškytė
- VU LSC - EMBL Partnership Institute For Gene Editing Technologies – Laboratory of Targeted CRISPR-Cas Delivery into Mammalian Nervous System, Lithuania
- Artūras Jakubauskas
- Hematology, Oncology and Transfusion Medicine Center, Vilnius University Hospital Santaros Klinikos, Vilnius, Lithuania
- Aurelijus Burokas
- Department of Biological Models, Institute of Biochemistry, Life Sciences Center, Vilnius University, Saulėtekio Ave. 7, LT-10257 Vilnius, Lithuania
- Ivan Nalvarte
- Karolinska Institutet, Department of Neurobiology, Care Sciences and Society, BioClinicum, Visionsgatan 4, Solna, Sweden
- Jose Inzunza
- Karolinska Institutet Stem Cell Organoid (KISCO) Facility, Department of Laboratory Medicine, Alfred Nobels allé 8, Huddinge, Sweden; Karolinska Institutet, Department of Laboratory Medicine, Alfred Nobels allé 8, Huddinge, Sweden
- Daniel Naumovas
- VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania; Laboratory of Nuclease Enabled Cell Therapies, Lithuania; Doctoral Program of Biology, Vilnius University Life Science Center, Lithuania; Hematology, Oncology and Transfusion Medicine Center, Vilnius University Hospital Santaros Klinikos, Vilnius, Lithuania
- Mindaugas Stoškus
- Hematology, Oncology and Transfusion Medicine Center, Vilnius University Hospital Santaros Klinikos, Vilnius, Lithuania
- Laimonas Griškevičius
- Hematology, Oncology and Transfusion Medicine Center, Vilnius University Hospital Santaros Klinikos, Vilnius, Lithuania
- Daiva Baltriukienė
- Department of Biological Models, Institute of Biochemistry, Life Sciences Center, Vilnius University, Saulėtekio Ave. 7, LT-10257 Vilnius, Lithuania
- Jonathan Arias
- VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania; Laboratory of Nuclease Enabled Cell Therapies, Lithuania; Karolinska Institutet Stem Cell Organoid (KISCO) Facility, Department of Laboratory Medicine, Alfred Nobels allé 8, Huddinge, Sweden; Karolinska Institutet, Department of Laboratory Medicine, Alfred Nobels allé 8, Huddinge, Sweden; Corresponding author at: VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania.
- Journal volume & issue
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Vol. 82
p. 103628
Abstract
Fluorescent protein-based Genetically Encoded Voltage Indicators (GEVI) offer a remarkable system for high-throughput screening of membrane potential phenotypes. The GEVI MARINA is a derivative from ArcLight, which conversely to ArcLight increases its fluorescence intensity alongside depolarization. Here we created knock-in reporter human iPS cell lines carrying the MARINA reporter using SpCas9 programmable nuclease and characterize a heterozygous clone.
