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Bruno Ngou @brunongou.bsky.social · 28d

Thank you Mary!!

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Bruno Ngou @brunongou.bsky.social · Sep 8

Thanks Nacho!

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Bruno Ngou @brunongou.bsky.social · Sep 5

Thank youuuu Pingtao

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Bruno Ngou @brunongou.bsky.social · Sep 5

Thank you Tatsuya!!

Bruno Ngou @brunongou.bsky.social · Sep 5

Thanks Adam!

Bruno Ngou @brunongou.bsky.social · Sep 4

I am very grateful to Michele & Marc, Takehiro & Dohmae-san, Markus, @yasukadota.bsky.social & @shirasulab.bsky.social for their support. Also big thanks to the reviewers and editors for improving our manuscript!

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Bruno Ngou @brunongou.bsky.social · Sep 4

With this approach, we aim to characterize more PRRs against different pathogens & pests in the future.

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Bruno Ngou @brunongou.bsky.social · Sep 4

To summarize, we reported the strategies to discover and engineer plant immune receptors.
As usual, we have uploaded the data, including RCM analysis, receptor & peptide sequences, raw data…etc etc on Zenodo
(zenodo.org/records/1546... )

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Bruno Ngou @brunongou.bsky.social · Sep 4

Together with structural prediction, phylogenomics data, and lots of in-planta validations, we engineered synthetic SCORE that perceive CSPs from a range of plant pathogens and pests.

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Bruno Ngou @brunongou.bsky.social · Sep 4

With the polymorphic CSP recognition specificity, we wonder if SCOREs can be engineered.
We predicted receptor-peptide structures to narrow down the region of interest.

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Bruno Ngou @brunongou.bsky.social · Sep 4

They do!
Across the 22 SOCRE orthologs we tested, most exhibit unique recognition specificity towards different CSP peptides.

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Bruno Ngou @brunongou.bsky.social · Sep 4

Bioinformatic analyses indicate that CSPs are very diverse across organisms (not just found in bacteria).
We wonder if SCORE orthologs across the flowering plant lineage can perceive different CSP peptides.

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Bruno Ngou @brunongou.bsky.social · Sep 4

Unlike the previously identified PRR CORE, "181" exhibits a more selective recognition towards csp15 variants, so we named it “Selective CORE” (SCORE).
In addition, CORE and SCORE are not closely related; these receptors convergently evolved to perceive CSPs.

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Bruno Ngou @brunongou.bsky.social · Sep 4

We screened for chimeric receptors activated by the plant pathogen Agrobacterium, and identified the receptor “181” from Pomelo.
Following biochemical analyses, we discovered that 181 perceives cold shock protein (CSP) peptides.

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Bruno Ngou @brunongou.bsky.social · Sep 4

To characterize immune receptors (PRRs) in plants, we cluster PRRs into subgroups based on conservation of their inner LRR residues.
We obtained >1000s subgroups with this approach, and cloned ~210s into receptor chimeras for further characterization.

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Bruno Ngou @brunongou.bsky.social · Sep 4

A🧵of the full story here:
bsky.app/profile/brun...

Below is an illustrative summary:

Bruno Ngou @brunongou.bsky.social · Mar 18

First post on Bluesky!! ☀️

Very happy to share that my main postdoc work ‘Systematic Discovery and Design of Synthetic Immune Receptors in Plants’ is finally out on Biorxiv!

www.biorxiv.org/content/10.1...

The full story is summarized here:

Systematic Discovery and Design of Synthetic Immune Receptors in Plants

Plants deploy a diverse array of pattern recognition receptors (PRRs) that perceive microbe-associated molecular patterns (MAMPs) to activate immune responses. Leucine-rich repeat receptor-like kinase...

www.biorxiv.org

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Bruno Ngou @brunongou.bsky.social · Sep 4

Very happy to share our latest work “Systematic discovery and engineering of synthetic immune receptors in plants” out in @science.org !

www.science.org/doi/10.1126/...

Systematic discovery and engineering of synthetic immune receptors in plants

Plants deploy a diverse array of pattern recognition receptors (PRRs), which perceive microbe-associated molecular patterns to activate immune responses. Leucine-rich repeat receptor-like kinase subgr...

www.science.org

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Bruno Ngou @brunongou.bsky.social · Mar 18

Yes! We are close to getting Arabidopsis transgenic lines that would recognize CSP from clubroot. Would be great to colab and test it!

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Bruno Ngou @brunongou.bsky.social · Mar 18

This has truly been one of my fav projects. Huge thanks to Michele & Marc, Takehiro & Dohmae-sensei for the bioinformatic & MS support, @yasukadota.bsky.social & @shirasulab.bsky.social for their unwavering support & belief in me & this project. Hope you enjoyed the thread & happy to discuss!

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Bruno Ngou @brunongou.bsky.social · Mar 18

To summarize, we:

-Designed a system to characterize the pathogen recognition landscape of LRR-RLK-XIIs in plants

-Identified an ancient but lineage-specific PRR that exhibits remarkable polymorphism in MAMP recognition

-Designed PRRs to perceive specific MAMPs from multiple crop pathogens

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Bruno Ngou @brunongou.bsky.social · Mar 18

Through this mini scan, we designed SCORE variants that robustly perceives CSPs from many devastating crop pathogens. Hopefully these would improve disease resistance against pathogens in crops
(XXI)

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Bruno Ngou @brunongou.bsky.social · Mar 18

We did a mini-mutational scan on these 3 AAs, generated 37 SCORE variants and tested them against 104 csp15 peptides. To a certain extent, the surface charges in this region determine csp15 recognition specificity in SCORE
(XX)

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Bruno Ngou @brunongou.bsky.social · Mar 18

Due to the AF prediction of ionic bonds between csp15 & 10th LRR of SCORE, we noticed that the surface charges in this region are different between SCORE orthologs. We can also drastically modify the surface charges, simply by mutating 3 AA residues in the 10th LRR
(XIX)

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Bruno Ngou @brunongou.bsky.social · Mar 18

This indicates that 8th-11th LRR determines csp15 recognition specificity in SCOREs. We perform a bunch of LRR swaps between orthologs and narrow it further down to the 10th LRR. csp15 recognition specificity can, in an extend, be transferred between SCORE orthologs
(XVIII)

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Bruno Ngou @brunongou.bsky.social · Mar 18

These predictions became convincing when combined with phylogenomics. While the AAs that interacts with csp15 in 3rd-7th & 12th-16thLRR are relatively conserved between SCORE orthologs, the LRR 8th-11th AAs that interacts with the variable region of csp15 are also variable
(XVII)

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