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| Status |
Public on Nov 15, 2015 |
| Title |
RNA Seq analysis of unchecked miR-998 expression in Drosophila melanogaster 3rd instar larval eye discs |
| Organism |
Drosophila melanogaster |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
The importance of the role of microRNAs in gene expression and disease is well recognized. However, what is less appreciated is that almost half of miRNA genes are organized in polycistronic clusters and are therefore co-expressed. The mir-11~998 cluster consists of two miRNAs, miR-11 and miR-998. Here, we describe a novel layer of regulation that links the processing and expression of miR-998 to the presence of the mir-11 gene. We show that the presence of mir-11 in the pri-miRNA is required for processing by Drosha, and deletion of mir-11 prevents the expression of miR-998. Replacing mir-11 with an unrelated miRNA rescued miR-998 expression in vivo and in vitro, as did expressing miR-998 from a shorter, more canonical miRNA scaffold. The embedded regulation of miR-998 is functionally important because unchecked miR-998 expression in the absence of miR-11 resulted in highly penetrant pleiotropic developmental defects. We further show that this novel regulation of expression of miRNAs within a cluster is not limited to the mir-11~998 cluster and likely reflects the more general cis-regulation of expression of individual miRNAs. Thus, our results reveal a novel layer of regulation within miRNA clusters that tempers the functions of the individual miRNAs. Unlinking their expression has the potential to change the expression of multiple miRNA targets and shift biological response.
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| Overall design |
RNA was extracted from Drosophila third instar larval eye discs of animals grown in standard conditions; Illumina HiSeq2000 Next Gen RNA Sequencing was performed, and differential expression of genes was assessed in wild-type vs unchecked miR-998 expression
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| Contributor(s) |
Truscott M, Islam AB, Frolov M |
| Citation(s) |
26554028 |
| |
| Submission date |
Oct 13, 2015 |
| Last update date |
Jun 07, 2019 |
| Contact name |
Mary Truscott |
| E-mail(s) |
[email protected]
|
| Organization name |
University of Illinois at Chicago
|
| Department |
Biochemistry and Molecular Genetics
|
| Lab |
Frolov lab
|
| Street address |
900 S Ashland Ave, room 2356
|
| City |
Chicago |
| State/province |
IL |
| ZIP/Postal code |
60607 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL13304 |
Illumina HiSeq 2000 (Drosophila melanogaster) |
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| Samples (8)
|
| GSM1906600 |
mir-11 KO(w-) biological replicate 2 |
| GSM1906601 |
mir-11~998 RMCE-1 biological replicate 1 |
| GSM1906602 |
mir-11~998 RMCE-1 biological replicate 2 |
| GSM1906603 |
mir-11~998 RMCE-2 biological replicate 1 |
| GSM1906604 |
mir-11~998 RMCE-2 biological replicate 2 |
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| Relations |
| BioProject |
PRJNA298583 |
| SRA |
SRP064742 |
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