Alice Ting

Life Science Research Professional 1 in School of Medicine, Stanford, California, United States We seek motivated recent college graduates with substantial research experience who are looking for a launching pad into the next phase of their...

subjects with at least 1 year of prior wetlab research experience. Must be a US citizen or have a valid work visa.

To apply: careersearch.stanford.edu/jobs/life-sc...

keeping the lab running (being a lab manager), and 50% is for original research related to protein engineering, technology development, etc – there are many project possibilities!

Seeking passionate, motivated, creative, and detail-oriented majors in chemical biology, molecular biology or related..

We are hiring a new lab manager! Our wonderful lab manager Albert is headed off to grad school and we are looking to fill his position this summer. This is a great opportunity for a new graduate to spend a couple years immersed in research before applying to PhD or MD programs. 50% of the job is...

Directed evolution of a sequence-specific covalent protein tag for RNA labeling | PNAS Efficient methods for conjugating proteins to RNA are needed for RNA delivery, imaging, editing, interactome mapping, and barcoding applications. N...

applications.

Congratulations to postdoc Rongbing Huang, whose creativity, persistence, and resourcefulness made rHUH possible! We are grateful to NSF and Biohub for funding this work.

www.pnas.org/doi/10.1073/pnas.2422085122

and imperfect sequence specificity resulting in significant background labeling of endogenous transcripts.

There is still work to do for this to be a dream tool for RNA imaging/editing/mapping in living cells, but rHUH provides a good starting point and it is definitely good enough for in vitro

library diversification.

The final rHUH tag is 13.4 kD, has 12 mutations relative to HUH, and covalently labels as little as 1 nM target RNA within minutes. The labeling works well in mammalian cell lysate, but not yet in the interior of living cells – possibly due to the high Mg2+ requirement

Today we report the directed evolution of HUH into rHUH, a sequence-specific covalent protein tag for RNA labeling. rHUH was hard to engineer: the initial HUH template had no RNA activity, so we started with RNA-DNA hybrids. 30 rounds of selection on the yeast surface were required, and 7 cycles of

There is no detectable toxicity in the dark state because the protease activity is completely shut off. After activation, LATeNT only cleaves VAMP2, inhibiting synaptic transmission, but it does not affect other pathways. It does not kill cells.

Yes we did a side by side comparison to PA-BoNT in Figure S7A and found that PA-BoNT showed much lower +light activity and higher dark-state background than LATeNT

Very proud of amazing protein engineer Heegwang Roh, our fantastic neuroscience collaborator Ji Won Um, co-first author Dongwook Kim from the Um lab, and our other wonderful co-authors.

Light-activated tetanus neurotoxin for conditional proteolysis and inducible synaptic inhibition in vivo The light chain of tetanus neurotoxin (TeNT) is a 52 kD metalloprotease that potently inhibits synaptic transmission by cleaving the endogenous vesicle fusion protein VAMP2. To mitigate the toxicity o...

effect are completely under your control.

Preprint here: www.biorxiv.org/content/10.1...

24 hours after shutting off LATeNT by taking away the light, anxiety levels returned to normal.

We are excited for neuroscientists to try this new tool! The inhibitory effect is stronger and more sustained than with other tools like halorhodopsin and DREADDs, but the timing and location of the...

Synaptic inhibition is potently inhibited only in those cells expressing LATeNT while neighboring cells are unaffected.

We used LATeNT in the mouse brain to discover a new cell population (SST+ interneurons in the hippocampus) that modulates anxiety.

we fused a light-sensitive LOV domain from oat plants into tetanus’ protease domain and made its activity controllable by light.

LATeNT (for light-activated tetanus neurotoxin) can be expressed in specific cell populations, and then turned on with a brief pulse of blue light.

Painting by Sir Charles Bell depicting opisthotonos, which can be caused by tetanus (1809)

Tetanus on demand! (not for bioweapons, but for neuroscience research)

Tetanus toxin induces paralysis by using a protease to cleave VAMP2, a protein required for synaptic vesicle fusion and neurotransmitter release.

To harness this toxicity and use it as a beneficial tool for neuroscience,

Global Science Scholars Postdoctoral Fellowship Program SS-F Apply for the new international postdoctoral fellowship program from the CZ Biohub Network and Stellar Science Foundation.

...research labs that are part of the CZ Biohub network (including mine!).

See czbiohub.org/program-ssfc... to learn more and email me with any questions

Applications accepted on a rolling basis until March 27, 2025

Global Science Scholars Postdoctoral Fellowship Program SS-F Apply for the new international postdoctoral fellowship program from the CZ Biohub Network and Stellar Science Foundation.

Are you a Japanese student interested in a postdoctoral fellowship in the US? The Chan Zuckerberg Initiative has an exciting new fully-funded Global Science Scholars program to pair students who have received their undergraduate or doctoral degrees in Japan with US bioengineering and biomedical...

activation migration, etc). This is discussed in the paper

CARs are much more restricted in both input scope (cell-attached antigens only) and output scope (immune activation only). PAGERs can respond to both soluble and cell-attached antigens and provide a wide range of outputs in response (transgene expression, real time fluorescence, inhibition,

Nick @nkalogriopoulos.bsky.social is on the job market this year and brimming over with creativity and deep experience. Nick did his PhD in GPCR structural bio. He is fearless, creative, rigorous, a great mentor and collaborator. His interests span synthetic bio, chem bio, and immunology.

This work was co-led by two extraordinary postdocs, @nkalogriopoulos.bsky.social l & @reikatei.bsky.social. We had wonderful collabs with yulonglilab.bsky.social, Yuqi Yan, Ivan Soltesz & Peter Klein. In our lab, Matt Ravalin helped characterize PAGERs, and Bo Cai assisted with macrophage expts.

Synthetic receptors such as CARs and synNotch have revolutionized cell therapies and provided powerful tools for basic research. We hope that PAGERs will contribute new capabilities to these exciting fields as well.

in response, leading to increased tumor cell killing. In macrophage cultures, PAGER-TF shifted macrophages to an M1 anti-tumor state, in the presence of tumor or M2 markers. Lastly, we expressed PAGER-Gi in human primary T cells to program them to migrate towards a soluble antigen gradient.

To demonstrate the utility of PAGER, we used it for antigen-control of neuronal excitation and inhibition, in both culture and acute brain slices. We also used PAGER in the cancer immunology context. PAGER-TF sensed tumor antigens and secreted an anti-CD3/anti-CD19 BiTE (Bispecific T cell Engager)

a real-time fluorescence response to antigen binding. And PAGER-G couples antigen recognition to activation of endogenous G protein pathways. Because there are four major classes of G proteins, we created four PAGER-G subtypes, for controlling endogenous Gi, Gs, Gq, or G12.

necessary. Potentially a wide array of binders can now be converted in a single step into receptors via the PAGER platform. Three different PAGER scaffolds provide access to distinct cellular responses to antigen activation. PAGER-TF converts antigen binding into transgene expression. PAGER-FL gives

We showed that PAGER is modular by inserting 15 different GFP or mCherry-binding nanobodies, as well as 13 nanobodies against growth factors, chemokines, cell surface receptors, etc., to create new antigen-activated synthetic receptors. No optimization, linker screening, or directed evolution was…

with the antagonist, leading to relief of receptor inhibition. Drug or agonist can then turn on PAGER.

www.nature.com/articles/s41...

which themselves are not structurally modular, but we were able to build in modular antigen gating by fusing an antagonist peptide to the extracellular N-terminal end, and then gating the antagonist with a fused antigen-binding nanobody. When antigen binds, it sterically interferes with the...

of innate cell behavior (excitation or inhibition of neurons, induction of cell migration, etc.)? Today we report in Nature a new platform that provides a step in that direction: PAGERs convert antigen recognition into diverse user-selected responses. PAGERs are based on G-protein coupled receptors