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Overexpressing Long Noncoding RNAs Using Gene-activating CRISPR

Article DOI: 10.3791/59233-v • 13:04 min • March 1st, 2019
March 1st, 2019 •
1, 1, 2, 1, 3, 3, 1, 1
1Vatche and Tamar Manoukian Division of Digestive Diseases, Department of Medicine, University of California, Los Angeles, 2Vatche and Tamar Manoukian Division of Digestive Diseases, Integrated Molecular Technologies (IMT) Core, Department of Medicine, University of California, Los Angeles, 3Division of Hematology and Oncology, Department of Medicine, University of California, Los Angeles

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Traditional cDNA-based overexpression techniques have a limited applicability for the overexpression of long noncoding RNAs due to their multiple splice forms with potential functionality. This review reports a protocol using CRISPR technology to overexpress multiple splice variants of a long noncoding RNA.

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Long Noncoding RNAs Gene-activating CRISPR Isoforms In Vitro Overexpression Studies Endogenous Isoforms Genome Expression Guide RNAs Gene-splicing Machinery Transcriptional Start Site Genetic Databases Deactivated Cas9 Plasmids E.coli Luria Broth Agar Plate Ampicillin LB (Luria-Bertani) Medium
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