Selection of anti-ARC antibodies (v1)

We exploited our Intracellular Antibodies Capture Technology (IACT) to select intrabodies against ARC (Activity-Regulated Cytoskeleton-associated protein) from a VHH Llama library in order to obtain ideal nanoscale tools to achieve ARC interference, imaging and even degradation in different subcellular compartments. Specifically, a conformer sensitive selection was designed against the NT-region (NT) of ARC (1-140), in which a 7 amino acids motif (113-119) has been demonstrated to be critical for oligomerization and capsid formation. Out of 80 initial candidates that were also re-screened against the NT-region in which the critical motif was mutated to 7 Alanine residues (s113-119a), we successfully identified 9 nanobodies (Nbs). These Nbs form part of a robust toolbox for targeting ARC protein with highly spatially-restricted subcellular localization (even in spines), allowing for modulation of function at those sites and interference with specific stages in the process from oligomerization to capsids. This dataset contains 9 anti-ARC Antibodies sequences (DNA/protein sequences and the predicted AlphaFold3 structure). The nanobody sequences provided can be used to express/produce the corresponding nanobodies to be employed either as intracellularly expressed antibodies to study ARC function in a cellular context or as proteins (imaging tools for ARC detection).