New dual-plasmid modifying system for DNA-based info rewriting in vivo

DNA-based info is a brand new interdisciplinary area linking info expertise and biotechnology. The sector hopes to satisfy the big want for long-term knowledge storage through the use of DNA as an info storage medium. Regardless of DNA’s promise of sturdy stability, excessive storage density and low upkeep price, nevertheless, researchers face issues precisely rewriting digital info encoded in DNA sequences.

Typically, DNA knowledge storage expertise has two modes, i.e., the “in vitro laborious disk mode” and the “in vivo CD mode.” The first benefit of the in vivo mode is its low-cost, dependable replication of chromosomal DNA by cell replication. Because of this attribute, it may be used for speedy and low-cost knowledge copy dissemination. Since encoded DNA sequences for some info include a lot of repeats and the looks of homopolymers, nevertheless, such info can solely be “written” and “learn,” however can’t be precisely “rewritten.”

To resolve the rewriting drawback, Prof. Liu Kai from the Division of Chemistry, Tsinghua College, Prof. Li Jingjing from the Changchun Institute of Utilized Chemistry (CIAC) of the Chinese language Academy of Sciences, and Prof. Chen Dong from Zhejiang College led a analysis group 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 device. This dual-plasmid system is appropriate for storing, studying and rewriting varied kinds of info, together with textual content, codebooks and pictures. It totally explores the coding functionality of DNA sequences with out requiring any addressing indices or backup sequences. It’s also 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 realize excessive effectivity in addition to reliability in rewriting advanced 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 might be addressed and rewritten. Due to 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. Because of optimizing the crRNA sequence, the data rewriting device turned extremely adaptable to advanced info, thus leading to rewriting reliability of as much as 94%, which is corresponding to present gene-editing techniques.

The twin-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 complex knowledge on a molecular stage.

We imagine this technique can be utilized in a dwelling host with a bigger genome, equivalent to yeast, which might additional pave the way in which for sensible functions relating to massive knowledge storage.”

Prof. Liu Kai, Division of Chemistry, Tsinghua College

Journal reference:

Liu, Y., et al. (2022) In vivo processing of digital info molecularly with focused specificity and strong reliability. Science Advances.



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