DNA-based Data Storage Research Articles

Optimizing fountain codes for DNA data storage

Fountain codes, originally developed for reliable multicasting in communication networks, are effectively applied in various data transmission and storage systems. Their recent use in DNA data storage systems has unique challenges, since the DNA storage channel deviates from the traditional Gaussian white noise erasure model considered in communication networks and has several restrictions as well as special properties. Thus, optimizing fountain codes to address these challenges promises to improve their overall usability in DNA data storage systems. In this article, we present several methods for optimizing fountain codes for DNA data storage. Apart from generally applicable optimizations for fountain codes, we propose optimization algorithms to create tailored distribution functions of fountain codes, which is novel in the context of DNA data storage. We evaluate the proposed methods in terms of various metrics related to the DNA storage channel. Our evaluation shows that optimizing fountain codes for DNA data storage can significantly enhance the reliability and capacity of DNA data storage systems. The developed methods represent a step forward in harnessing the full potential of fountain codes for DNA-based data storage applications.The new coding schemes and all developed methods are available under a free and open-source software license.

A Cost-Effective Approach for Single-Stranded DNA Amplification Using Primer-Blocked Asymmetric PCR.

In vitro amplification of single-stranded oligonucleotide libraries presents a significant challenge due to the potential for excessive byproduct formation. This phenomenon largely affects the quality of the ssDNAs created using the most commonly used methods, e.g., asymmetric PCR, biotin-streptavidin separation, or lambda exonuclease digestion of dsDNA. Here, we describe an improved protocol that combines primer-blocked asymmetric PCR (PBA-PCR) with emulsion PCR and a cost-effective downstream process that altogether alleviates byproduct formation without distorting the sequence space of the ssDNA library. In PBA-PCR, the reaction mixture is complemented with a 3'-phosphate-blocked limiting primer that decreases mispriming, thus reducing polymerization of DNA byproducts. The downstream process includes mixing of the PBA-PCR product with excess reverse complement of the 3'-phosphate-blocked limiting primer and removal of dsDNA strands via biotin-streptavidin separation, yielding purified ssDNAs. In conclusion, we have devised a universally applicable approach for simple and cost-effective production of ssDNA libraries and unique ssDNA sequences with on-demand labeling. Our protocol could be beneficial for a variety of uses, such as generating aptamer libraries for SELEX, creating unique molecular identifiers for a wide range of sequencing applications, providing donor DNA for CRISPR-Cas9 systems, developing scaffold nanostructures, and enabling DNA-based data storage. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Amplification of ssDNA libraries using PBA-PCR Alternate Protocol 1: Amplification of ssDNA libraries using emulsion PBA-PCR with a simplified extraction of PBA-PCR products Basic Protocol 2: Purification of PBA-PCR products to remove dsDNA and conversion of 3'-blocked primer to double-stranded complexes Alternate Protocol 2: Purification of PBA-PCR products to remove both dsDNA and blocking primers from the reaction mixture Support Protocol: Analysis of PBA-PCR products by gel electrophoresis.

“Multi-layer” encryption of medical data in DNA for highly-secure storage

The exponential increasement and the attributes of medical data drive the requirement for secure medical data archiving. DNA data storage shows promise for storing sensitive and important data like medical records due to its high density and endurance. Nevertheless, current DNA data storage working scheme generally does not fully consider the data encryption, posing a risk of data corruption by routine DNA sequencing. Here, we designed a “multi-layer” encryption pipeline for medical data archiving. Initially, digital information was encrypted using Blowfish algorithm at information technology (IT) layer, followed by two-layer data encryption at the biotechnology (BT) layer. The first BT layer exploited the molecular weight of synthetic DNA or nucleoside to encrypt the key, while the second BT layer encrypted digital information within DNA sequences. Consequently, decryption involved layer-by-layer interpretation of data, including mass spectroscopy, sequencing, and Blowfish decryption, significantly enhancing data security. Utilizing mass spectroscopy to retrieve information allows for employment of both natural and unnatural nucleosides, as well as their synthetic oligonucleotides, for data storage, thereby considerably boosting scalability. Our work implies expanded flexibility of DNA-based data storage, highlighting the potential for leveraging various physical and chemical characteristics of DNA molecules to encode and access digital information.

Toward DNA-Based Recording of Biological Processes.

Exploiting the inherent compatibility of DNA-based data storage with living cells, various cellular recording approaches have been developed for recording and retrieving biologically relevant signals in otherwise inaccessible locations, such as inside the body. This review provides an overview of the current state of engineered cellular memory systems, highlighting their design principles, advantages, and limitations. We examine various technologies, including CRISPR-Cas systems, recombinases, retrons, and DNA methylation, that enable these recording systems. Additionally, we discuss potential strategies for improving recording accuracy, scalability, and durability to address current limitations in the field. This emerging modality of biological measurement will be key to gaining novel insights into diverse biological processes and fostering the development of various biotechnological applications, from environmental sensing to disease monitoring and beyond.

Explorer: efficient DNA coding by De Bruijn graph toward arbitrary local and global biochemical constraints.

With the exponential growth of digital data, there is a pressing need for innovative storage media and techniques. DNA molecules, due to their stability, storage capacity, and density, offer a promising solution for information storage. However, DNA storage also faces numerous challenges, such as complex biochemical constraints and encoding efficiency. This paper presents Explorer, a high-efficiency DNA coding algorithm based on the De Bruijn graph, which leverages its capability to characterize local sequences. Explorer enables coding under various biochemical constraints, such as homopolymers, GC content, and undesired motifs. This paper also introduces Codeformer, a fast decoding algorithm based on the transformer architecture, to further enhance decoding efficiency. Numerical experiments indicate that, compared with other advanced algorithms, Explorer not only achieves stable encoding and decoding under various biochemical constraints but also increases the encoding efficiency and bit rate by ¿10%. Additionally, Codeformer demonstrates the ability to efficiently decode large quantities of DNA sequences. Under different parameter settings, its decoding efficiency exceeds that of traditional algorithms by more than two-fold. When Codeformer is combined with Reed-Solomon code, its decoding accuracy exceeds 99%, making it a good choice for high-speed decoding applications. These advancements are expected to contribute to the development of DNA-based data storage systems and the broader exploration of DNA as a novel information storage medium.

Information Density Enhancement Using Lossy Compression in DNA Data Storage.

This study develops two deoxyribonucleic acid (DNA)lossy compression models, Models A and B, to encode grayscale images into DNA sequences, enhance information density, and enable high-fidelity image recovery. These models, distinguished by their handling of pixel domains and interpolation methods, offer a novel approach to data storage for DNA. Model A processes pixels in overlapped domains using linear interpolation (LI), whereas Model B uses non-overlapped domains with nearest-neighbor interpolation (NNI). Through a comparative analysis with Joint Photographic Experts Group (JPEG)compression, the DNA lossy compression models demonstrate competitive advantages in terms of information density and image quality restoration. The application of these models to the Modified National Institute of Standards and Technology (MNIST) dataset reveals their efficiency and the recognizability of decompressed images, which is validated by convolutional neural network (CNN) performance. In particular, Model B2, a version of Model B, emerges as an effective method for balancing high information density (surpassing over 20 times the typical densities of two bits per nucleotide) with reasonably good image quality. These findings highlight the potential of DNA-based data storage systems for high-density and efficient compression, indicating a promising future for biological data storage solutions.

Guest Editorial Plenty of Room at at Bottom: Ten Years of DNA-Based Data Storage

Guest Editorial Plenty of Room at at Bottom: Ten Years of DNA-Based Data Storage

An Effective DNA-Based File Storage System for Practical Archiving and Retrieval of Medical MRI Data.

DNA-based data storage is a new technology in computational and synthetic biology, that offers a solution for long-term, high-density data archiving. Given the critical importance of medical data in advancing human health, there is a growing interest in developing an effective medical data storage system based on DNA. Data integrity, accuracy, reliability, and efficient retrieval are all significant concerns. Therefore, this study proposes an Effective DNA Storage (EDS) approach for archiving medical MRI data. The EDS approach incorporates three key components (i) a novel fraction strategy to address the critical issue of rotating encoding, which often leads to data loss due to single base error propagation; (ii) a novel rule-based quaternary transcoding method that satisfies bio-constraints and ensure reliable mapping; and (iii) an indexing technique designed to simplify random search and access. The effectiveness of this approach is validated through computer simulations and biological experiments, confirming its practicality. The EDS approach outperforms existing methods, providing superior control over bio-constraints and reducing computational time. The results and code provided in this study open new avenues for practical DNA storage of medical MRI data, offering promising prospects for the future of medical data archiving and retrieval.

Data recovery methods for DNA storage based on fountain codes

Today's digital data storage systems typically offer advanced data recovery solutions to address the problem of catastrophic data loss, such as software-based disk sector analysis or physical-level data retrieval methods for conventional hard disk drives. However, DNA-based data storage currently relies solely on the inherent error correction properties of the methods used to encode digital data into strands of DNA. Any error that cannot be corrected utilizing the redundancy added by DNA encoding methods results in permanent data loss. To provide data recovery for DNA storage systems, we present a method to automatically reconstruct corrupted or missing data stored in DNA using fountain codes. Our method exploits the relationships between packets encoded with fountain codes to identify and rectify corrupted or lost data. Furthermore, we present file type-specific and content-based data recovery methods for three file types, illustrating how a fusion of fountain encoding-specific redundancy and knowledge about the data can effectively recover information in a corrupted DNA storage system, both in an automatic and in a guided manual manner. To demonstrate our approach, we introduce DR4DNA, a software toolkit that contains all methods presented. We evaluate DR4DNA using both in-silico and in-vitro experiments.

Towards the development of an electrochemical random access DNA memory (e-RADM)

As a result of the exponentially growing amount of information being produced, new data storage solutions are required. DNA has attracted significant attention as a potential data storage medium thanks to several intrinsic properties, such as ultrahigh physical information density with up to 2 bits per nucleotide. Current DNA-based memories rely on sequencing strategies for data recovery. However, sequencing all DNA strands for data retrieval would be very time consuming and thus result in high levels of latency. Therefore, random access strategies are required to make DNA-based data storage a viable alternative. Here, we present our first steps towards the development of a compartmentalized electrochemical random access DNA memory (e-RADM) using cascade reactions controlled by DNA nanostructures immobilized on gold microelectrode arrays which will be triggered when a specific information retrieval query is put into the system. Electrodes containing the desired information can then be identified by Square Wave Voltammetry.Graphical abstract

Efficient DNA-based data storage using shortmer combinatorial encoding

Data storage in DNA has recently emerged as a promising archival solution, offering space-efficient and long-lasting digital storage solutions. Recent studies suggest leveraging the inherent redundancy of synthesis and sequencing technologies by using composite DNA alphabets. A major challenge of this approach involves the noisy inference process, obstructing large composite alphabets. This paper introduces a novel approach for DNA-based data storage, offering, in some implementations, a 6.5-fold increase in logical density over standard DNA-based storage systems, with near-zero reconstruction error. Combinatorial DNA encoding uses a set of clearly distinguishable DNA shortmers to construct large combinatorial alphabets, where each letter consists of a subset of shortmers. We formally define various combinatorial encoding schemes and investigate their theoretical properties. These include information density and reconstruction probabilities, as well as required synthesis and sequencing multiplicities. We then propose an end-to-end design for a combinatorial DNA-based data storage system, including encoding schemes, two-dimensional (2D) error correction codes, and reconstruction algorithms, under different error regimes. We performed simulations and show, for example, that the use of 2D Reed-Solomon error correction has significantly improved reconstruction rates. We validated our approach by constructing two combinatorial sequences using Gibson assembly, imitating a 4-cycle combinatorial synthesis process. We confirmed the successful reconstruction, and established the robustness of our approach for different error types. Subsampling experiments supported the important role of sampling rate and its effect on the overall performance. Our work demonstrates the potential of combinatorial shortmer encoding for DNA-based data storage and describes some theoretical research questions and technical challenges. Combining combinatorial principles with error-correcting strategies, and investing in the development of DNA synthesis technologies that efficiently support combinatorial synthesis, can pave the way to efficient, error-resilient DNA-based storage solutions.

Concatenated Nanopore DNA Codes.

In nanopore sequencers, single-stranded DNA molecules (or k-mers) enter a small opening in a membrane called a nanopore and modulate the ionic current through the pore, producing a channel output in the form of a noisy piecewise constant signal. An important problem in DNA-based data storage is finding a set of k-mers, i.e. a DNA code, that is robust against noisy sample duplication introduced by nanopore sequencers. Good DNA codes should contain as many k-mers as possible that produce distinguishable current signals (squiggles) as measured by the sequencer. The dissimilarity between squiggles can be estimated using a bound on their pairwise error probability, which is used as a metric for code design. Unfortunately, code construction using the union bound is limited to small k's due to the difficulty of finding maximum cliques in large graphs. In this paper, we construct large codes by concatenating codewords from a base code, thereby packing more information in a single strand while retaining the storage efficiency of the base code. To facilitate decoding, we include a circumfix in the base code to reduce the effect of the nanopore channel memory. We show that the decoding complexity scales as [Formula: see text], where m is the number of concatenated k-mers. Simulations show that the base code error rate is stable as m increases.

DNA Bloom Filter enables anti-contamination and file version control for DNA-based data storage.

DNA storage is one of the most promising ways for future information storage due to its high data storage density, durable storage time and low maintenance cost. However, errors are inevitable during synthesizing, storing and sequencing. Currently, many error correction algorithms have been developed to ensure accurate information retrieval, but they will decrease storage density or increase computing complexity. Here, we apply the Bloom Filter, a space-efficient probabilistic data structure, to DNA storage to achieve the anti-error, or anti-contamination function. This method only needs the original correct DNA sequences (referred to as target sequences) to produce a corresponding data structure, which will filter out almost all the incorrect sequences (referred to as non-target sequences) during sequencing data analysis. Experimental results demonstrate the universal and efficient filtering capabilities of our method. Furthermore, we employ the Counting Bloom Filter to achieve the file version control function, which significantly reduces synthesis costs when modifying DNA-form files. To achieve cost-efficient file version control function, a modified system based on yin-yang codec is developed.

FrameD: Framework for DNA-based Data Storage Design, Verification, and Validation.

DNA-based data storage is a quickly growing field that hopes to harness the massive theoretical information density of DNA molecules to produce a competitive next-generation storage medium suitable for archival data. In recent years, many DNA-based storage system designs have been proposed. Given that no common infrastructure exists for simulating these storage systems, comparing many different designs along with many different error models is increasingly difficult. To address this challenge we introduce FrameD, a simulation infrastructure for DNA storage systems that leverages the underlying modularity of DNA storage system designs to provide a framework to express different designs while being able to reuse common components. We demonstrate the utility of FrameD and the need for a common simulation platform using a case study. Our case study compares designs that utilize strand copies differently, some that align strand copies using Multiple Sequence Alignment (MSA) algorithms and others that do not. We found that the choice to include MSA in the pipeline is dependent on the error rate and the type of errors being injected and is not always beneficial. In addition to supporting a wide range of designs, FrameD provides the user with transparent parallelism to deal with a large number of reads from sequencing and the need for many fault injection iterations. We believe that FrameD fills a void in the tools publicly available to the DNA storage community by providing a modular and extensible framework with support for massive parallelism. As a result, it will help accelerate the design process of future DNA-based storage systems. The source code for FrameD along with the data generated during the demonstration of FrameD is available in a public Github repository at https://github.com/dna-storage/framed (10.5281/zenodo.7757762).

Reducing cost in DNA-based data storage by sequence analysis-aided soft information decoding of variable-length reads.

DNA-based data storage is one of the most attractive research areas for future archival storage. However, it faces the problems of high writing and reading costs for practical use. There have been many efforts to resolve this problem, but existing schemes are not fully suitable for DNA-based data storage, and more cost reduction is needed. We propose whole encoding and decoding procedures for DNA storage. The encoding procedure consists of a carefully designed single low-density parity-check code as an inter-oligo code, which corrects errors and dropouts efficiently. We apply new clustering and alignment methods that operate on variable-length reads to aid the decoding performance. We use edit distance and quality scores during the sequence analysis-aided decoding procedure, which can discard abnormal reads and utilize high-quality soft information. We store 548.83 KB of an image file in DNA oligos and achieve a writing cost reduction of 7.46% and a significant reading cost reduction of 26.57% and 19.41% compared with the two previous works. Data and codes for all the algorithms proposed in this study are available at: https://github.com/sjpark0905/DNA-LDPC-codes.

The Secure Storage Capacity of a DNA Wiretap Channel Model

In this paper, we propose a strategy for making DNA-based data storage information-theoretically secure through the use of wiretap channel coding. This motivates us to extend the shuffling-sampling channel model of Shomorony and Heckel (2021) to include a wiretapper. Our main result is a characterization of the <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">secure storage capacity</i> of our DNA wiretap channel model, which is the maximum rate at which data can be stored within a pool of DNA molecules so as to be reliably retrieved by an authorized party (Bob), while ensuring that an unauthorized party (Eve) gets almost no information from her observations. Furthermore, our proof of achievability shows that index-based wiretap channel coding schemes are optimal.

A Segmented-Edit Error-Correcting Code With Re-Synchronization Function for DNA-Based Storage Systems

As a powerful tool for storing digital information in chemically synthesized molecules, DNA-based data storage has undergone continuous development and received increasingly more attention. Efficiently recovering information from large-scale DNA strands that suffer from insertions, deletions, and substitution errors (collectively referred to as edit errors), is one of the major bottlenecks in DNA-based storage systems. To cope with this challenge, in this paper, we provide a segmented-edit error-correcting code with the re-synchronization function, termed the <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">DNA-LM</i> code. Compared with the previous segmented-error-correcting codes, it has a systematic structure and does not require the endpoint of the received segment as pre-requisite information for decoding. In the case that the number of edit errors exceeds the edit error-correcting capability of a segment, it can easily regain synchronization to ensure that the subsequent decoding continues. Both encoding and decoding complexity is linear in the codeword length. The redundancy of each segment is <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex-math notation="LaTeX">$\lceil \log k\rceil +6$</tex-math></inline-formula> quaternary symbols, where <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex-math notation="LaTeX">$k$</tex-math></inline-formula> is the length of the message segment. We further generalize the decoding algorithm to deal with duplicated DNA strands, whereas it still maintains linear time complexity in the codeword length and the number of duplications. Simulations under a stochastic edit errors model show that, at a low raw error rate of the “next-gen” sequencing, our code can enable error-free decoding by concatenating with the (255,223) RS code.

Towards long double-stranded chains and robust DNA-based data storage using the random code system.

DNA has become a popular choice for next-generation storage media due to its high storage density and stability. As the storage medium of life's information, DNA has significant storage capacity and low-cost, low-power replication and transcription capabilities. However, utilizing long double-stranded DNA for storage can introduce unstable factors that make it difficult to meet the constraints of biological systems. To address this challenge, we have designed a highly robust coding scheme called the "random code system," inspired by the idea of fountain codes. The random code system includes the establishment of a random matrix, Gaussian preprocessing, and random equilibrium. Compared to Luby transform codes (LT codes), random code (RC) has better robustness and recovery ability of lost information. In biological experiments, we successfully stored 29,390 bits of data in 25,700 bp chains, achieving a storage density of 1.78 bits per nucleotide. These results demonstrate the potential for using long double-stranded DNA and the random code system for robust DNA-based data storage.

Integrating FPGA Acceleration in the DNAssim Framework for Faster DNA-Based Data Storage Simulations

DNA-based data storage emerged in this decade as a promising solution for long data durability, low power consumption, and high density. However, such technology has not yet reached a good maturity level, requiring many investigations to improve the information encoding and decoding processes. Simulations can be key to overcoming the time and the cost burdens of the many experiments imposed by thorough design space explorations. In response to this, we have developed a DNA storage simulator (DNAssim) that allows simulating the different steps in the DNA storage pipeline using a proprietary software infrastructure written in Python/C language. Among the many operations performed by the tool, the edit distance calculation used during clustering operations has been identified as the most computationally intensive task in software, thus calling for hardware acceleration. In this work, we demonstrate the integration in the DNAssim framework of a dedicated FPGA hardware accelerator based on the Xilinx VC707 evaluation kit to boost edit distance calculations by up to 11 times with respect to a pure software approach. This materializes in a clustering simulation latency reduction of up to 5.5 times and paves the way for future scale-out DNA storage simulation platforms.

Basis-Finding Algorithm for Decoding Fountain Codes for DNA-Based Data Storage

In this paper, we consider the decoding of fountain codes where the received symbols may have errors. It is motivated by the application of fountain codes in DNA-based data storage systems where the inner code decoding, which generally has undetectable errors, is performed before the outer fountain code decoding. We propose a novel and efficient decoding algorithm, namely basis-finding algorithm (BFA), followed by three implementations. The key idea of the BFA is to find a basis of the received symbols, and then use the most reliable basis elements to recover the source symbols with the inactivation decoding. Gaussian elimination is used to find the basis and to identify the most reliable basis elements. As a result, the BFA has polynomial time complexity. For random fountain codes, we are able to derive some theoretical bounds for the frame error rate (FER) of the BFA. Extensive simulations with Luby transform (LT) codes show that, the BFA has significantly lower FER than the belief propagation (BP) algorithm except for an extremely large amount of received symbols, and the FER of the BFA generally decreases as the average weight of basis elements increases.