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rank phylum # of entries
1 Ascomycota 4629 (79%)
2 Basidiomycota 1116 (19%)
3 Mucoromycotina 38 (1%)
Development of DNA sequence tagging tools based on machine learning using
public sequence annotation data [配列注釈オープンデータに基づくDNA Smart Taggerの開発] Eli Kaminuma1, Takatomo Fujisawa1, Fumi Hayashi1, Tatsuya Nishizawa2, Yasukazu Nakamura1, Osamu Ogasawara1
(1. Center for Information Biology, National Institute of Genetics, 2. IMSBIO Co., Ltd. )
ABSTRACT Large-scale DNA sequence data generated from New Generation Sequencers (NGSs) is published from the International Nucleotide Sequence Database Collaboration (INSDC) in addition to
journal reports. Large scale open data of DNA sequences and the annotations could be basic data materials of machine learning models. However applications of large-scale DNA database to machine
learning models are not enough to be developed for practical many situations. We suppose that sequence registration tools to be submitted to DNA data banks enable to support users to select appropriate
annotation attributes by machine learning models. In this research, we aim to construct DNA Smart Tagging tools, where the tool name is “DNASmartTagger”, that predict candidate attribute values of
annotation tags for DNA sequences. As the first step, we define the methodology of parametrization strategy for DNA sequence-based machine learning models. Moreover model performances and application
domains of respective annotation tags are discussed. Practically, in experiment the 4,850 sequences of 16S rRNA gene in INSDC BCT division were retrieved to train supervised machine learning models for
/PCR_primers annotation tag. In a similar procedure, the 5,831 fungal sequences of INSDC PLN division were utilized for constructing predictive models for /altitude annotation tag. Support Vector Machine
(SVM), a supervised learning method, was used for classifying attribute values of individual tags. For evaluating models, the area under the ROC curve (AUC) of SVMs based on 5’end short fragment inputs
was computed. AUC for /PCR_primers tag and /altitude tag were 0.83 and 0.79 individually, where fragment lengths were 60bp and 128bp. Further, the SVMs for the /altitude tag exhibited 0.87 AUC when
input parameter was normalized frequency of k-mer indices. In detail, the input parameter means normalized frequency of appearance of possible 5-mers over not short fragment but full-length sequence.
The result may indicate that k-mer based approach can work more efficiently than short fragment based approach.
Acknowledgments: ・ We would like to thank Dr. Jun Mashima, Koji Watanabe, Prof. Toshihisa Takagi for constructive comments and technical assistance.
・ This work was partially supported by CREST, JST and ROIS management expenses grant.
・ Computations were partially performed on the NIG supercomputer at ROIS National Institute of Genetics.
Publication:
• DNA Data Bank of Japan. Mashima J, Kodama Y, Fujisawa T, Katayama T, Okuda Y, Kaminuma E, Ogasawara O, Okubo K, Nakamura Y, Takagi T.Nucleic Acids Res. 2017 45:D25-D31.
• The International Nucleotide Sequence Database Collaboration. Cochrane G, Karsch-Mizrachi I, Takagi T, International Nucleotide Sequence Database Collaboration, Nucleic Acids Res. 2016 44:D48-50.
• DDBJ new system and service refactoring. Ogasawara O, Mashima J, Kodama Y, Kaminuma E, Nakamura Y, Okubo K, Takagi T. Nucleic Acids Res. 2013 41:D25-9.
Result(1) predictive performance of classification models for /PCR_primers attribute values
The problem for high labor costs of manual annotation at sequence submission stage to DNA data bank
Experimental conditions (preparing training datasets)
■ Investigating Predictive Performances by Input Types • Study type( whole genome sequencing, metagenomics, amplicon sequencing, etc ) • Indirect encoding(without responsive loci) vs direct encoding with responsive loci • Whole genome markers vs fragment markers
■ Evaluating Performances for Other Tags by Heterogeneous Divisions
e.g. /altitude contained divisions (bacteria, virus, etc.)
■ Designing the DNASmartTagger API System (under development) • Model retraining function with successively new data release • Visualizing multiple tag candidates
Future Work
Background:Next-Generation DNA Sequencing Produces Large Quantities of Data
Problem: Detailed Manual Annotations Lead to High Labor Costs
# of Released Entries
DBCLS SRA statistics (Nakazato et al., 2013)
http://sra.dbcls.jp/
DDBJ Trad DDBJ SRA (NGS Raw
Reads)
2016 196M 91K
2015 189M 62K
Accacactggtactgagacacgga
ccagactcctacgggaggcagcag
tgaggaatattggacaatggaggga
actctgatccagccatgccgcgtgca
ggaagactgccctatgggttgtaaac
tgcttttatacaagaagaataagaga
tacgtgtatcttgatgacggtattgtaa
gaataagcaccggctaactccgtgc
cagcagccgcggtaatacggaggg
tgcaagcgttatccggaatcattgggt
ttaaagggtccgtaggcggattaata
agtcagtggtgaaagtctgcagctta
actgtagaattgccattgatactgtta
gtcttgaattattatgaagtagttagaa
tatgtagtgtagcggtgaaatgcata
gatattaca
Input: DNA Sequence
sequence
e.g. INSDC FlatFile Format
Altitudinal zonation
Output: Annotation Tags
DNASmartTagger
Utilizing open data
for training models BioSample
452 attribute tags INSDC 89 qualifier key tags
Machine Learning Models
GBIF, etc.
DDBJ ANNOTATION HELP
DNASmartTagger : a proposed machine learning method for predicting structured DNA sequence annotation
Annotations
Result(2) predictive performances of classification models for /altitude attribute values
/altitude INSDC Qualifier Tag /PCR_primers INSDC Qualifier Tag
/altitude tag
■ Predicting target keys for pilot studies = INSDC qualifier keys (/altitude, /PCR_primers)
* Sequence annotation of SRA BioSample contains many unstructured contents requiring exhaustive data cleansing.
* INSDC sequence annotations are well controlled compared to SRA BioSample annotations.
■ Sequences and annotation data were retrieved from the DDBJ ARSA data retrieval tool.
TAG Output
Variable Type
# of
Entries
ML Model Design Classification Performance
(AUC wt Cross-Validation)
Data Retrieval
Condition
1 /PCR_Primers
Categorical
(Multilabel)
4,850
Support Vector
Machine(SVM)
5’end fragment
(L=60bp)
0.83 [37 PrimerFwd models]
0.81 [104 PrimerRev models]
BCT Division
16S rRNA
TAG Output Variable
Type
# of
Entries
ML Model
Design
Classification Performance
(AUC wt Cross-Validation)
Data Retrieval
Condition
2 /altitude Continuous
↓※
Categorical
(Multilabel)
5,831 SVM, 3 models
5’end fragment (L=128bp)
0.79
[Z1=0.81, Z2=0.73, Z3=0.83 ]
- PLN Division
- Fungi
(keyword)
5-mers frequency for full-length
(※)
0.87
[Z1=0.91, Z2=0.80, Z3=0.91 ]
acagagttttcggactgctg
acgaccggcgcacgggtg
cgtaacgcgtatacaatcta
cc
acagagttttcggactgctgacgaccggcgcacgggtgcgtaacgcgtatacaatctaccttttgctaa
gggatagcccagagaaatttggattaatactttatggtatgtatttatggcatcatatatacattaaaggtt
acggcaaaagatgagtatgcgttctattagctagatggtaaggtaacggcttaccatggctacgatag
ataggggccctgagagggggatcccccacactggtactgagacacggaccagactcctacggga
ggcagcagtgaggaatattggacaatggagggaactctgatccagccatgccgcgtgcaggaaga
ctgccctatgggttgtaaactgcttttatacaagaagaataagagatacgtgtatcttgatgacggtattg
taagaataagcaccggctaactccgtgccagcagccgcggtaatacggagggtgcaagcgttatcc
ggaatcattgggtttaaagggtccgtaggcggattaataagtcagtggtgaaagtctgcagcttaactg
tagaattgccattgatactgttagtcttgaattattatgaagtagttagaatatgtagtgtagcggtgaaat
gcatagatattacatagaataccgattgcgaaggcaggctactaataatatattgacgctgatggacg
aaagcgtgggtagcgaacaggattagataccctggtagtccacgccgtaaacgatggtcactagct
gttcggacttcggtctgagtggctaagcgaaagtgataagtgacccacctggggagtacgttcgcaa
gaatgaaact
16S rRNA sequences (5’3’) 5’end fragment
agtggctaagcgaaagtga
taagtgacccacctgggga
gtacgttcgcaagaatgaa
act
PrimerFWD candidate:F27 (agagtttgatcmtggctcag)
PrimerREV candidate:1525R (aaggaggtgwtccarcc)
3’end fragment
■ Model Performance
■ Fragmentation processing of input sequence for /PCR_Primers tag prediction
■ Frequency of entries by
sequence length
# o
f entr
ies b
y m
odels
■ Model Performance
ZONE Attribute Value Altitude Zone Code
ALPINE ZONE 1500m -- Z3
MONTANE ZONE 800m--1500m Z2
LOWLAND ZONE 0--800m Z1
AU
C
■ Length of /altitude retrieved sequences
Unique taxonmy ID.= 3,667
Unique attribute value = 257
Unique PrimerFWD seq. = 107
Unique PrimerREV seq,.= 115
■ Classification performance with frequency of entries
by PrimerFWD sequence [fragment length=60]
■ Classification performance
by input fragment length (training dataset)
fragment length [# of entries ≧ 20] AUC=Area Under the Curve, 37 PrimerFwd models [# of entries ≧ 20]
rank primer set target
loci
# of
entries
1 ITS1 - ITS4 ITS 228
2 ITS5 - ITS4 ITS 103
3 ITS5 - NL4 ITS, LSU 93
4 nu-ssu-0817 –
nu-ssu-1536
SSU 76
5 niaD15F - niaD12R euknr 21
■Top5 primer set (annotated only)
■Top3 phyla
※Ref. Fiannaca et al., A k-mer-based barcode DNA classification methodology , AI in Medicine 64:173, 2015
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