Neuron-Specific Genome Modification in the Adult Rat Brain Using CRISPR-Cas9 Transgenic Rats

Neuron
Susanne BäckBrandon K Harvey

Abstract

Historically, the rat has been the preferred animal model for behavioral studies. Limitations in genome modification have, however, caused a lag in their use compared to the bevy of available transgenic mice. Here, we have developed several transgenic tools, including viral vectors and transgenic rats, for targeted genome modification in specific adult rat neurons using CRISPR-Cas9 technology. Starting from wild-type rats, knockout of tyrosine hydroxylase was achieved with adeno-associated viral (AAV) vectors expressing Cas9 or guide RNAs (gRNAs). We subsequently created an AAV vector for Cre-dependent gRNA expression as well as three new transgenic rat lines to specifically target CRISPR-Cas9 components to dopaminergic neurons. One rat represents the first knockin rat model made by germline gene targeting in spermatogonial stem cells. The rats described herein serve as a versatile platform for making cell-specific and sequence-specific genome modifications in the adult brain and potentially other Cre-expressing tissues of the rat.

Citations

Dec 6, 2019·International Journal of Molecular Sciences·Julia KonovalovaAndrii Domanskyi
Sep 5, 2019·International Journal of Molecular Sciences·Shao-Cheng WangChing-Ming Cheng
Jun 7, 2019·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·Kristina HerfertBernd J Pichler
Jul 25, 2020·Nature Reviews. Neuroscience·Yun Young YimEric J Nestler
Mar 12, 2020·Animal Cells and Systems·Hyunji LeeKyoungmi Kim
Sep 19, 2020·American Journal of Physiology. Heart and Circulatory Physiology·Satoshi KobaTatsuo Watanabe
Dec 15, 2020·Biomaterials Science·João ConniotJoão Conde
Dec 22, 2020·Synthetic Biology·Grace A MeakerThomas E Gorochowski
Feb 23, 2021·The European Journal of Neuroscience·Cecilia P KramarM Flavia Barbano

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