Targeted Deletion of Zebrafish lncRNAis18 with TALENs
dc.contributor.author | Jones, Crystal | |
dc.contributor.department | Genetics, Development and Cell Biology | |
dc.date | 2018-02-14T10:03:36.000 | |
dc.date.accessioned | 2020-07-07T05:10:45Z | |
dc.date.available | 2020-07-07T05:10:45Z | |
dc.date.issued | 2014-04-15 | |
dc.description.abstract | <p>Long non-coding RNAs (lncRNAs) are important players in epigenetic regulation of gene expression during development and disease (Niland et al, 2012). A number of mechanisms have been proposed for lncRNA action, however, few functional studies of lncRNAs have been described. We are using Transcription Activator-Like Effector Nucleases (TALENs), engineered site-specific nucleases, to create targeted mutations in a novel zebrafish lncRNA. We previously mapped a highly penetrant retinal tumor model to transgene disruption of the zebrafish lncRNAis18 gene. The objective of this project is to isolate a second zebrafish lncRNAis18 allele that contains a deletion of part of the lncRNAis18 gene. Two TALEN pairs were designed to simultaneously target double-strand breaks to exons 2 and 5 of lncRNAis18. Injection of 25-40pg of the TALENs targeting individual exons into zebrafish embryos resulted in efficient mutagenesis of the target sites. To isolate the lncRNAis18 deletion allele we co-injected embryos with the TALEN pairs targeting both exons 2 and exon 5. We predicted co-injection of TALEN pairs targeting exons 2 and 5 of lncRNAis18 would create a 147kb deletion after loss of the intervening sequence and repair by the non-homologous enjoining pathway. PCR products spanning the fusion of exons 2 to 5 were amplified from somatic tissue in 9 out of 14 co-injected embryos. We verified the deletion allele by sequencing PCR products from 3 embryos. We have identified one founder that transmits the deletion allele to the F1 generation. F1 embryos are being raised to establish a new lncRNAis18del line. The lncRNAis18 deletion allele will provide a new genetic tool to study the function of lncRNAis18 in zebrafish development and cancer.</p> | |
dc.identifier | archive/lib.dr.iastate.edu/undergradresearch_symposium/2014/presentations/70/ | |
dc.identifier.articleid | 1069 | |
dc.identifier.contextkey | 5914631 | |
dc.identifier.s3bucket | isulib-bepress-aws-west | |
dc.identifier.submissionpath | undergradresearch_symposium/2014/presentations/70 | |
dc.identifier.uri | https://dr.lib.iastate.edu/handle/20.500.12876/91637 | |
dc.relation.ispartofseries | Symposium on Undergraduate Research and Creative Expression | |
dc.source.bitstream | archive/lib.dr.iastate.edu/undergradresearch_symposium/2014/presentations/70/JonesCrystal.pdf|||Sat Jan 15 01:40:09 UTC 2022 | |
dc.subject.disciplines | Genetics | |
dc.title | Targeted Deletion of Zebrafish lncRNAis18 with TALENs | |
dc.type | event | |
dc.type.genre | article | |
dspace.entity.type | Publication | |
relation.isOrgUnitOfPublication | 9e603b30-6443-4b8e-aff5-57de4a7e4cb2 | |
relation.isSeriesOfPublication | 6730f354-97b8-4408-bad3-7e5c3b2fca9d | |
thesis.degree.discipline | Genetics |
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