Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/84342
Title: Ribozyme Mediated gRNA Generation for In Vitro and In Vivo CRISPR/Cas9 Mutagenesis
Authors: Lee, Raymond Teck Ho
Ng, Ashley Shu Mei
Ingham, Philip William
Keywords: Embryos
Ribozymes
Issue Date: 2016
Source: Lee, R. T. H., Ng, A. S. M., & Ingham, P. W. (2016). Ribozyme Mediated gRNA Generation for In Vitro and In Vivo CRISPR/Cas9 Mutagenesis. PLOS ONE, 11(11), e0166020-.
Series/Report no.: PLOS ONE
Abstract: CRISPR/Cas9 is now regularly used for targeted mutagenesis in a wide variety of systems. Here we report the use of ribozymes for the generation of gRNAs both in vitro and in zebrafish embryos. We show that incorporation of ribozymes increases the types of promoters and number of target sites available for mutagenesis without compromising mutagenesis efficiency. We have tested this by comparing the efficiency of mutagenesis of gRNA constructs with and without ribozymes and also generated a transgenic zebrafish expressing gRNA using a heat shock promoter (RNA polymerase II-dependent promoter) that was able to induce mutagenesis of its target. Our method provides a streamlined approach to test gRNA efficiency as well as increasing the versatility of conditional gene knock out in zebrafish.
URI: https://hdl.handle.net/10356/84342
http://hdl.handle.net/10220/41788
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0166020
Rights: © 2016 Lee et al. This is an open access article distributed underthe terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:LKCMedicine Journal Articles

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