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

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

5,283 Views

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11:24 min

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May 14, 2019

DOI:

10.3791/59578-v

DOI:
10.3791/59578-v

•

11:24 min
May 14, 2019

•

4 Views
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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

Summary

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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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LentiviralGene SilencingHuman PseudoisletLow Attachment PlatesIslet FunctionGene Expression ModulationSHR8 TransductionProteolytic EnzymeCollagenolytic EnzymeCell DissociationCMRL MediumFetal Bovine Serum

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Cite this Article

Liu, S., Harata, M., Promes, J. A., Burand, A. J., Ankrum, J. A., Imai, Y. Lentiviral Mediated Gene Silencing in Human Pseudoislet Prepared in Low Attachment Plates. J. Vis. Exp. (147), e59578, doi:10.3791/59578 (2019).

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