期刊
RNA
卷 22, 期 8, 页码 1120-1138出版社
COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1261/rna.055509.115
关键词
small RNA sequencing; microRNA; next-generation sequencing; short-read aligners; gene expression
资金
- Juvenile Diabetes Research Foundation International (JDRF)
- National Health and Medical Research Council (NHMRC)
- National Heart Foundation of Australia (NHF)
- NHMRC
- Victorian Government's Operational Infrastructure Support Program
Genomic alignment of small RNA (smRNA) sequences such as microRNAs poses considerable challenges due to their short length (similar to 21 nucleotides [nt]) as well as the large size and complexity of plant and animal genomes. While several tools have been developed for high-throughput mapping of longer mRNA-seq reads (>30 nt), there are few that are specifically designed for mapping of smRNA reads including microRNAs. The accuracy of these mappers has not been systematically determined in the case of smRNA-seq. In addition, it is unknown whether these aligners accurately map smRNA reads containing sequence errors and polymorphisms. By using simulated read sets, we determine the alignment sensitivity and accuracy of 16 short-read mappers and quantify their robustness to mismatches, indels, and nontemplated nucleotide additions. These were explored in the context of a plant genome (Oryza sativa, similar to 500 Mbp) and a mammalian genome (Homo sapiens, similar to 3.1 Gbp). Analysis of simulated and real smRNA-seq data demonstrates that mapper selection impacts differential expression results and interpretation. These results will inform on best practice for smRNA mapping and enable more accurate smRNA detection and quantification of expression and RNA editing.
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