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Hello @micronaut.bsky.social. Some of our gRNAs are indeed targeting either of the strand but we did not observe black and white differences. However, the set of gRNAs is too small to draw a definite conclusion. I suggest trying nicking either of the strands if this is a concern for your phage :)

That would be amazing indeed 😀

We did not test this but I would cautiously say, not necessarily. If phage SSBs (or other repair proteins) are recruited to fix the nick, then it could work in other phages as well. What would be interesting is to know how phages fix ssDNA breaks and how universal it is.

Whilst we suspect repair is Gp2.5 or SSB related, it is difficult to prove as these proteins are essential for phage viability. Also, these proteins interact with the polymerase so I would doubt if they can be used universally.

Many thanks to Frank Englert, Sarshad K. Valappil, Jonas Kubilius, @drjonesky.bsky.social , @vivekmutalik.bsky.social , and Chase Beisel. #Bacteriophage #PhageEngineering #GenomeEditing #CRISPR #PhageTherapy

4/4 We then used this library to isolate T7 variants capable of infecting otherwise resistant, LPS-deficient E. coli hosts, showing how scalable phage genome editing can be used to expand host range.

3/4 Maintaining high phage titers also makes the method well suited for large-scale mutagenesis. Using nCas9, we generated a library of more than 440,000 T7 tail-fiber variants in a single editing step.

2/4 We unexpectedly found that Cas9 nickase, which introduces only a single-strand DNA break, can edit selected phage genomes with remarkably high efficiency while preserving phage titers. Unlike many existing approaches, this avoids multi-step workflows, counterselection, and extensive screening.

🔗 Discover more: bit.ly/3LUzM2U #EMBO #GenomeEditing #CRISPR #SyntheticBiology #Microbiology #Biotechnology #Vilnius #LifeSciences

Dr C. Patinios Awarded EMBO Installation Grant to Advance Genome Editing Research in LithuaniaVilnius University is a prestigious institution of science and studies in Lithuania, which develops world-class science and develops science-based international studies.bit.ly

This work will drive new genome editing technologies and uncover fundamental DNA repair mechanisms, with impact across microbiome research, synthetic biology, industrial biotechnology, and healthcare.

🧬 My project, “Pause-Repair-Edit,” aims to move beyond current CRISPR tools by developing safer genome editing approaches that pause DNA replication to activate natural repair pathways—opening genome editing to many microorganisms that are currently hard to engineer.

Thank you Jens

Thank you Nicholas 😊

Amazing Martin!! All the best with the new lab and the amazing research you do!!!

Big shout-out to first authors Darshana Gupta, (myself :) ) & Harris V. Bassett, senior author Chase L. Beisel, and project initiator Scott P. Collins 🙌

This opens up brand-new possibilities for precision editing and studying DNA repair!

We developed append editing, a genome-editing strategy that installs ADP-ribose groups onto DNA to unlock new repair outcomes: 🔹 In bacteria → precise, scar-free edits via homologous recombination 🔹 In eukaryotes → unique T→A or T→C base substitutions not possible with current editors

Learn more ➡️ www.gmc.vu.lt/en/lsc-embl/...

The Laboratory of Dr. Constantinos PatiniosVU Gyvybės mokslų centras jungia tris VU padalinius – Biomokslų, Biotechnologijos ir Biochemijos institutus. Centras įkurtas vystant integruotą mokslo, studijų ir verslo centrą (slėnį) „Saulėtekis“.www.gmc.vu.lt

Deadline: Open until a suitable candidate is found. 📧 Send your CV + Motivation Letter to: constantinos.patinios@gmc.vu.lt

🧬 Our work focuses on next-generation genome editing tools for bacteria, yeasts, and phages — with a special focus on DNA replication stalling and repair!