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Genetics

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JoVE Journal Genetics
Lentiviral Mediated Gene Silencing in Human Pseudoislet Prepared in Low Attachment Plates
 

Lentiviral Mediated Gene Silencing in Human Pseudoislet Prepared in Low Attachment Plates

Article DOI: 10.3791/59578-v • 11:24 min • May 14th, 2019
May 14th, 2019 •
1,2, 1,2, 1,2, 2,3, 2,3, 1,2
1Department of Internal Medicine, Carver College of Medicine, University of Iowa, 2Fraternal Order of Eagles Diabetes Research Center, University of Iowa, 3Roy J. Carver Department of Biomedical Engineering, University of Iowa

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résumé
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A protocol to create gene modified human pseudoislets from dispersed human islet cells that are transduced by lentivirus carrying short hairpin RNA (shRNA) is presented. This protocol utilizes readily available enzyme and culture vessels, can be performed easily, and produces genetically modified human pseudoislets suitable for functional and morphological studies.

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Keywords: Lentiviral Gene Silencing Human Pseudoislet Low Attachment Plates Islet Function Gene Expression Modulation SHR8 Transduction Proteolytic Enzyme Collagenolytic Enzyme Cell Dissociation CMRL Medium Fetal Bovine Serum
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