Abstract
MicroRNAs are small 18–24 nt single-stranded noncoding RNA molecules involved in many biological processes, including stemness maintenance and cellular reprogramming. Current methods used in loss-of-function studies of microRNAs have several limitations. Here, we describe a new approach for dissecting miR-302/367 functions by transcription activator-like effectors (TALEs), which are natural effector proteins secreted by Xanthomonas and Ralstonia bacteria. Knockdown of the miR-302/367 cluster uses the Kruppel-associated box repressor domain fused with specific TALEs designed to bind the miR-302/367 cluster promoter. Knockout of the miR-302/367 cluster uses two pairs of TALE nucleases (TALENs) to delete the miR-302/367 cluster in human primary cells. Together, both TALE-based transcriptional repressor and TALENs are two promising approaches for loss-of-function studies of microRNA cluster in human primary cells.
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Zhang, Z., Wu, WS. (2018). Application of TALE-Based Approach for Dissecting Functional MicroRNA-302/367 in Cellular Reprogramming. In: Ying, SY. (eds) MicroRNA Protocols . Methods in Molecular Biology, vol 1733. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7601-0_21
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DOI: https://doi.org/10.1007/978-1-4939-7601-0_21
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7601-0
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