Category: DeNSAS

DeNSAS: De Novo Sequence Annotation System

Overview

DeNSAS (De Novo Sequence Annotation System) is a comprehensive, reference-free pipeline designed for functional annotation of transcripts, proteins, and genes in genome assemblies that lack reference annotation. Developed in-house, this automated system integrates multiple public databases and computational tools to provide accurate functional characterization of sequences without requiring a reference genome.

Core Functionality

DeNSAS employs a 6-step process using custom Perl scripts and locally built databases to provide:

  • Sequence Description Assignment: Integrating results from multiple database searches to assign comprehensive functional descriptions to sequences
  • Gene Ontology (GO) Annotation: Assignment of GO terms for biological process, molecular function, and cellular component
  • Protein Domain Identification: Recognition of conserved protein domains using PFAM
  • RNA Family Classification: Identification of RNA families using RFAM
  • Protease Classification: Specialized annotation of peptidases using MEROPS
  • Gene Family Assignment: Characterization of gene families based on sequence similarity

Key Features

  • Reference-Free Annotation: Functions independently without requiring a reference genome or annotation
  • Integrative Approach: Combines data from multiple databases including PFAM, RFAM, UniProt, InterPro, MEROPS, and RefSeq
  • Novel Gene Identification: Capable of characterizing potentially novel genes and duplication events
  • Comprehensive Output: Provides detailed annotation including:
    • Full gene names and unique IDs (when close matches exist)
    • Protein domains and motifs
    • Gene family assignments
    • GO term classifications
    • RNA family patterns

Advantages Over Existing Tools

  • Unlike reference-based tools (e.g., OrthoFinder, CRB-Blast), DeNSAS does not require an existing reference strain to guide annotation
  • Compared to other reference-free annotation pipelines (e.g., PANNZER2), DeNSAS offers improved accuracy through its integrative nature and multi-database approach
  • Custom filtering thresholds ensure high-quality annotations with minimal false positives

Technical Implementation

DeNSAS employs stringent similarity thresholds:

  • BLAST searches use an e-value cut-off of 10e⁻⁵ and HSP similarity threshold of 80%
  • Sequence descriptions employ more restrictive thresholds (e-value 1e-10 and 90% similarity)
  • RNA and protein family identification filtered by Expectation Value (1e-10)
  • Transparent labeling of sequences with insufficient database matches as "UNKNOWN DESCRIPTION"

Applications

  • Annotation of non-model organisms
  • Characterization of newly sequenced genomes
  • Identification of novel genes and gene families
  • Comparative genomic analyses
  • Functional studies of transcriptomes

Releases