Our proof of principle work relied on imaging test cargo proteins (e.g., GFP) for which cognate Darpin sequences had already been published. There are many [>100] known Darpin sequences selected for binding diverse targets , but our applications to DOE-mission-relevant targets requires development of an in-house pipeline for selecting Darpins for binding specific targets. In that effort, so far we have created a Darpin library (with variable loops) and piloted a bacterial-display syste
To reach higher resolution, we will use next-generation scaffolds we have recently developed (unpublished) in which the protruding Darpin appendages are rigidified by bridging interactions.
Using newer scaffolds under development, we hope to break the 3 A resolution barrier for the first time. If this is possible it will set another landmark for imaging small proteins by EM, with far-reaching impact.
Success in individual projects will advance our atomic understanding in specific areas related to cellulosic formation/degradation. This will impact the progress of internal institute projects (in the Microbe and Plant areas) as well as projects in collaborating BER funded laboratories (CCRC and UCSB).