Nontoxic, double-deletion-mutant rabies viral vectors for retrograde targeting of projection neurons

Soumya Chatterjee, Heather A. Sullivan, Bryan J. MacLennan, Ran Xu, YuanYuan Hou, Thomas K. Lavin, Nicholas E. Lea, Jacob E. Michalski, Kelsey R. Babcock, Stephan Dietrich, Gillian A. Matthews, Anna Beyeler, Gwendolyn G. Calhoon, Gordon Glober, Jennifer D. Whitesell, Shenqin Yao, Ali Cetin, Julie A. Harris, Hongkui Zeng, Kay M. Tye, R. Clay Reid, Ian R. Wickersham
Nat Neurosci. 2018-03-05; 21(4): 638-646
DOI: 10.1038/s41593-018-0091-7

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Recombinant rabies viral vectors have proven useful for applications including
retrograde targeting of projection neurons and monosynaptic tracing, but their
cytotoxicity has limited their use to short-term experiments. Here we introduce a
new class of double-deletion-mutant rabies viral vectors that left transduced
cells alive and healthy indefinitely. Deletion of the viral polymerase gene
abolished cytotoxicity and reduced transgene expression to trace levels but left
vectors still able to retrogradely infect projection neurons and express
recombinases, allowing downstream expression of other transgene products such as
fluorophores and calcium indicators. The morphology of retrogradely targeted
cells appeared unperturbed at 1 year postinjection. Whole-cell patch-clamp
recordings showed no physiological abnormalities at 8 weeks. Longitudinal
two-photon structural and functional imaging in vivo, tracking thousands of
individual neurons for up to 4 months, showed that transduced neurons did not die
but retained stable visual response properties even at the longest time points
imaged.

Auteurs Bordeaux Neurocampus