TRIPinRNA

Python RNA Biology Bioinformatics Gene Regulation X Chromosome Inactivation

Project Overview

TRIPinRNA is an innovative in silico platform for predicting intramolecular RNA triple helix structures. Triple helices are non-canonical RNA structures that play crucial roles in gene regulation, including X chromosome inactivation, genomic imprinting, and telomere maintenance. However, identifying and studying these structures experimentally is challenging and resource-intensive.

As lead developer, I created this computational tool to predict potential triple helices in RNA sequences using a sophisticated algorithm that evaluates sequence complementarity, structural constraints, and thermodynamic properties. The platform provides researchers with a valuable resource for investigating the functional roles of RNA triple helices in various biological contexts.

Triple Helix Prediction

Advanced algorithms to identify potential triple helix forming sequences in RNA

Thermodynamic Analysis

Evaluation of stability and formation likelihood based on thermodynamic parameters

Structural Visualization

Interactive visualization of predicted triple helix structures and their features

Comprehensive Database

Repository of known and predicted triple helix structures across various organisms

Research Impact

TRIPinRNA has made significant contributions to the field of RNA biology by providing new insights into the prevalence and roles of triple helices in gene regulation. Our publication in Biochemistry demonstrates the tool's capabilities and its applications in understanding complex regulatory mechanisms.

X Chromosome Inactivation

Identified potential triple helix structures in XIST RNA that may play roles in silencing the X chromosome. These predictions provide new avenues for experimental validation and deeper understanding of the molecular mechanisms involved in dosage compensation.

Gene Regulation Mechanisms

Discovered triple helix structures in the 3' UTRs of various genes, suggesting roles in post-transcriptional regulation. These findings contribute to our understanding of how RNA structure influences gene expression and cellular function.

Methodological Advances

Developed novel computational approaches for predicting RNA triple helices, addressing limitations in existing methods. The algorithms incorporate sequence complementarity, structural constraints, and thermodynamic considerations.

Experimental Validation

Successfully validated several predicted triple helices using biophysical techniques, demonstrating the accuracy and reliability of the TRIPinRNA platform. These validations provide confidence in the tool's predictions.

Technical Approach

The development of TRIPinRNA involved several key technical components and algorithms:

1

Sequence Complementarity Analysis

Algorithm to identify potential triplex-forming sequences based on Hoogsteen and Watson-Crick base-pairing rules. This step evaluates the potential for three RNA strands to interact and form stable structures.

2

Structural Constraint Evaluation

Assessment of structural feasibility based on constraints like loop length, distance between interacting regions, and steric considerations. These constraints ensure that the predicted structures can physically form within the RNA molecule.

3

Thermodynamic Stability Calculation

Implementation of energy models to evaluate the stability of predicted triple helices, incorporating nearest-neighbor parameters and environmental factors. This step assesses whether the structures are energetically favorable.

4

Machine Learning Classification

Development of a classifier to distinguish between true positives and false positives based on features extracted from known triple helices. This approach improves prediction accuracy by learning from verified examples.

5

Visualization Pipeline

Creation of a visualization system to represent predicted triple helices and their features in an intuitive and informative manner. The visualizations help researchers understand the structural details and potential biological significance.

Key Findings

Triple Helix Prevalence

Discovered that triple helices are more prevalent in the human transcriptome than previously thought, with particular enrichment in long non-coding RNAs involved in gene regulation.

Functional Associations

Identified associations between triple helix structures and specific functional roles, including chromatin interactions, RNA-protein binding, and RNA stability regulation.

Evolutionary Conservation

Observed significant evolutionary conservation of triple helix forming sequences across mammalian species, suggesting important functional roles that have been preserved.

Novel RNA Motifs

Discovered novel sequence motifs associated with triple helix formation that were not previously characterized in the literature, expanding our understanding of RNA structures.

Publication

Development of an In Silico Platform (TRIPinRNA) for the Identification of Novel RNA Intramolecular Triple Helices and Their Validation Using Biophysical Techniques

Isha Rakheja*, Vishal Bharti*, Souvik Maiti, and Debojyoti Chakraborty

Biochemistry (2024)

Abstract: RNA molecules can form a variety of secondary and tertiary structures that play important roles in cellular processes. One such structure is the triple helix, where a third strand interacts with a duplex through Hoogsteen or reverse Hoogsteen base pairing. In this study, we present TRIPinRNA, a computational platform for predicting intramolecular RNA triple helices. The platform incorporates algorithms for sequence complementarity analysis, structural constraint evaluation, and thermodynamic stability calculation. We validate our predictions using biophysical techniques including circular dichroism spectroscopy and thermal melting experiments. Our findings reveal that triple helices are more prevalent in the human transcriptome than previously recognized, with particular enrichment in long non-coding RNAs involved in gene regulation. This platform provides researchers with a valuable resource for investigating the functional roles of RNA triple helices in various biological contexts.

Future Directions

The development of TRIPinRNA opens up several exciting avenues for future research:

Enhanced Prediction Algorithms

Continuing to refine the prediction algorithms by incorporating deep learning approaches and additional experimental data. These improvements will enhance the accuracy and reliability of triple helix predictions.

Expanded Database

Building a comprehensive database of RNA triple helices across diverse organisms, providing a valuable resource for comparative studies and evolutionary analyses. This database will facilitate the identification of conserved structural patterns.

Integration with Other Tools

Developing integrations with other RNA structure prediction tools to create a more comprehensive platform for RNA structural analysis. This integration will provide researchers with a more complete picture of RNA folding and structure.

Experimental Validation Pipeline

Creating a systematic pipeline for experimental validation of predicted triple helices, including protocols for in vitro and cellular assays. This pipeline will help bridge the gap between computational predictions and biological reality.

Interested in RNA Structure Research?

If you're interested in learning more about TRIPinRNA, collaborating on RNA structure research, or discussing potential applications, I'd be happy to connect.