New dual-plasmid modifying system for DNA-based info rewriting in vivo
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DNA-based info is a brand new interdisciplinary subject linking info know-how and biotechnology. The subject hopes to satisfy the big want for long-term knowledge storage by utilizing DNA as an info storage medium. Despite DNA’s promise of sturdy stability, excessive storage density and low upkeep price, nonetheless, researchers face issues precisely rewriting digital info encoded in DNA sequences.

Generally, DNA knowledge storage know-how has two modes, i.e., the “in vitro hard disk mode” and the “in vivo CD mode.” The major benefit of the in vivo mode is its low-cost, dependable replication of chromosomal DNA by cell replication. Due to this attribute, it may be used for fast and low-cost knowledge copy dissemination. Since encoded DNA sequences for some info include numerous repeats and the looks of homopolymers, nonetheless, such info can solely be “written” and “read,” however can’t be precisely “rewritten.”

To resolve the rewriting drawback, Prof. Liu Kai from the Department of Chemistry, Tsinghua University, Prof. Li Jingjing from the Changchun Institute of Applied Chemistry (CIAC) of the Chinese Academy of Sciences, and Prof. Chen Dong from Zhejiang University led a analysis crew that not too long ago developed a dual-plasmid modifying system for precisely processing digital info in a microbial vector. Their findings had been printed in Science Advances.

The researchers established a dual-plasmid system in vivo utilizing a rationally designed coding algorithm and an info modifying software. This dual-plasmid system is appropriate for storing, studying and rewriting varied forms of info, together with textual content, codebooks and pictures. It absolutely explores the coding functionality of DNA sequences with out requiring any addressing indices or backup sequences. It can be suitable with varied sorts of coding algorithms, thus enabling excessive coding effectivity. For instance, the coding effectivity of the present system reaches 4.0 bits per nucleotide.

To obtain excessive effectivity in addition to reliability in rewriting complicated info saved in exogenous DNA sequences in vivo, a wide range of CRISPR-associated proteins (Cas) and recombinase had been used. The instruments had been guided by their corresponding CRISPR RNA (crRNA) to cleave a goal locus in a DNA sequence in order that the precise info may very well be addressed and rewritten. Because of the excessive specificity between complementary pairs of nucleic acid molecules, the information-encoded DNA sequences had been precisely reconstructed by recombinase to encode new info. Due to optimizing the crRNA sequence, the data rewriting software turned extremely adaptable to complicated info, thus leading to rewriting reliability of as much as 94%, which is similar to current gene-editing programs.

The dual-plasmid system can function a common platform for DNA-based info rewriting in vivo, thus providing a brand new technique for info processing and target-specific rewriting of enormous and sophisticated knowledge on a molecular degree.

We consider this technique can be utilized in a residing host with a bigger genome, equivalent to yeast, which might additional pave the best way for sensible purposes relating to large knowledge storage.”

Prof. Liu Kai, Department of Chemistry, Tsinghua University

Source:

Journal reference:

Liu, Y., et al. (2022) In vivo processing of digital info molecularly with focused specificity and sturdy reliability. Science Advances. doi.org/10.1126/sciadv.abo7415.

Posted in: Genomics

Tags: Biotechnology, Cell, CRISPR, DNA, Gene, Gene-Editing, Genome, in vitro, in vivo, Locus, Nucleic Acid, Nucleotide, Plasmid, Research, RNA, Yeast

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