Tag: TnpB

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Cryo-EM structures of the D. radiodurans ISDra2 TnpB binary and ternary complexes. Credit: Nature (2023). DOI: 10.1038/s41586-023-05826-x A team led by Professor Virginijus Šikšnys from Vilnius University Life Sciences Center (VU LSC) determined the structure of TnpB using cryo-electron microscopy in collaboration with the group of Professor Guillermo Montoya at…

Register for FREE to listen to this article Thank you. Listen to this article using the player above. ✖ Want to listen to this article for FREE? Complete the form below to unlock access to ALL audio articles. Gene editing is one of the latest breakthroughs in…

TnpB adopts a bilobed structure consisting of the REC lobe and NUC lobe. The REC lobe comprises WED and REC domains, whereas the NUC lobe comprises the RuvC and TNB domains. TnpB assembles with a ωRNA transcribed from the transposon sequence at the end of its coding region. The guide…

Sample preparation The genes encoding ISDra2 TnpB (TnpB from D. radiodurans ISDra2; residues 1 to 408) and the 247-nt ωRNA were synthesized by Eurofin Genomics and cloned into the modified pETDuet vector (Novagen). The N-terminally MBP-tagged TnpB and ωRNA were co-expressed in Escherichia coli Rosetta2 (DE3). The E. coli cells were cultured…

Summary January 20th, 2023 A protocol is outlined to perform live real-time imaging to quantify how the accessory protein TnpB affects the dynamics of transposition in individual live Escherichia coli cells. Transcript This protocol allows us to quantify excision statistics on a single-cell basis, as opposed to looking at bulk…

20 September 2021 Researchers looking for evolutionary ancestors of enzymes used in the CRISPR genome editing have uncovered over one million potential genome-editing enzymes. The CRISPR/Cas9 approach uses small pieces of RNA as guides to direct a DNA-cutting enzyme called Cas9 to edit specific locations in the genome. It is this…

By analyzing microbial genomes, researchers have found what may be evolutionary ancestors of the Cas9 enzyme that snips out bits of DNA during CRISPR genome editing, according to a study published last week in Science.   According to Nature, the role of the microbial IscB protein family was unknown prior to the…

Thanks to CRISPR, gene therapy and “designer babies” are now a reality. The gene editing Swiss army knife is one of the most impactful biomedical discoveries of the last decade. Now a new study suggests we’ve just begun dipping our toes into the CRISPR pond. CRISPR-Cas9 comes from lowly origins….

While investigating the ancestry of Cas proteins, the researchers also found the first evidence of CRISPR activity in a eukaryotic cell. CRISPR gene editing is a field with many promises that has sometimes struggled to deliver. Now, a new study published in the journal Science has made a step towards…

Researchers seeking to understand the origins of CRISPR-Cas9 systems have uncovered a class of transposon-encoded RNA-guided nucleases, which they dubbed “OMEGA,” that could be harnessed for genome editing in human cells. These systems hold strong potential for developing as biotechnologies, the authors say. CRISPR-Cas systems have transformed genome editing and…

Left: Han Altae-Tran (Credit: Zhang Lab). Right: Soumya Kannan. Credit: Caitlin Cunningham Within the last decade, scientists have adapted CRISPR systems from microbes into gene editing technology, a precise and programmable system for modifying DNA. Now, scientists at MIT’s McGovern Institute and the Broad Institute of MIT and Harvard have…

The Cas9 enzyme — known as CRISPR’s molecular scissors — is used to find and cut specific DNA sequences.Credit: Nature By exploring the evolutionary origins of an enzyme used in the CRISPR genome-editing system, researchers have unearthed more than one million other potential editors lurking in microbial genomes. The study,…

NEW YORK – A team of US researchers led by the Broad Institute’s Feng Zhang has discovered a new class of transposon-encoded RNA-guided DNA nucleases, which they said could be used for genome editing in human cells and hold potential for biotechnology. In a paper published on Thursday in Science,…