TINY RNA, BIG DREAMS: HOW BRIDGE RNA SHAKING UP GENE EDITING
In the ever-changing field of genetic engineering, genetically designed products and technologies are essential. This recent work reveals the mechanism of action of bridge RNAs as a gene editing tool. According to earlier research, enzymes have a variety of functions in the opportunistic spread of viruses, the synthesis of new genes, and mobile genetic elements (MGE). MGEs are DNA segments that are mobile inside and across genomes without relying on host replication.
The nc-RNA (non-coding RNA) was discovered to be a crucial component for
the recombination after the researchers' earlier attempt at IS110 invitro
recombination failed. IS110’s circular nc-RNA structures feature two internal
loops, serving by bridge RNA to both target and donor DNA. The circularization
of the IS110 to nc-RNA activates the recombinase. This enzyme consists of
serine residues in the C-terminal and DEDD (group amino acids) in the
N-terminal. From the study it was understood that IS621 recombinase from the
IS110 family had a higher affinity for both the donor and the target,
which led to selecting it as potential gene editing approach. It was noted that the donor DNA attached to
the second terminal loop while the target DNA bound to the first. The flexible
nature of the bridge RNA was the researcher's main focus. It's interesting to
note that by modifying the sequence surrounding its core nucleotide sequence,
the bridge RNA permits precise targeting.
Figure
1: Base-pairing model of the IS621 bridge RNA with cognate target and donor DNA.
Combined analysis of
Covariation analysis and Base-pairing concordance analysis indicated potential
base-pairing between nc-RNA and the target and donor. It is also inferred that
the 5’ side of each loop binds with bottom strand of the target or donor with a
stretch of 8 nucleotides whereas 3’ side of each loop binds with the top strand
of the target or donor using 4 to 7 nucleotides.
Researchers conducted a comprehensive study on bridge RNA and IS621 genomic insertion, employing diverse oligo pools to assess donor-binding loop programmability and efficiency across guide sequences. Using advanced sequencing techniques like long-read nanopore sequencing, they identified precise insertion junctions in the E. coli genome, evaluating insertion specificity and off-target effects. Computational pipelines detected structural variants and analyzed target specificity, revealing insights into bridge RNA function and genomic interactions. These findings highlight bridge RNA's potential in genetic engineering, emphasizing its role in precise genome editing strategies.
In conclusion, the groundbreaking findings of this study open up exciting new possibilities in genetic engineering. The potential to revolutionize gene editing in various biological system is immerse. As researcher dwell deep in to this field, they may uncover similar insertion sequence with similar function. The integration of bridge RNA with advanced techniques like nanopore could enable rapid and accurate assigning editing efficiency and specificity. This leap forward of new era of precision and effectiveness in genetic engineering
Reference:
Durrant MG, Perry NT, Pai JJ, Jangid AR, Athukoralage JS, Hiraizumi M,
McSpedon JP, Pawluk A, Nishimasu H, Konermann S, Hsu PD. Bridge RNAs direct
programmable recombination of target and donor DNA. Nature. 2024 Jun
27;630(8018):984-93.
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