@article{602a4fc35353439f8877460c71e3a23c,
title = "Neuron-Specific Genome Modification in the Adult Rat Brain Using CRISPR-Cas9 Transgenic Rats",
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. Limited availability of transgenic rats has hindered their utility in modeling human behavior and cognition. B{\"a}ck, Necarsulmer, et al. describe new transgenic rats and viral vectors that enable neuron- and region-specific gene modifications within the adult rat brain.",
keywords = "AAV, CRISPR-Cas9, MANF, brain, genome editing, lsl-Cas9, spermatogonial stem cells, transgenic rat",
author = "Susanne B{\"a}ck and Julie Necarsulmer and Whitaker, {Leslie R.} and Coke, {Lamarque M.} and Pyry Koivula and Heathward, {Emily J.} and Fortuno, {Lowella V.} and Yajun Zhang and Yeh, {C. Grace} and Baldwin, {Heather A.} and Spencer, {Morgan D.} and Mejias-Aponte, {Carlos A.} and James Pickel and Hoffman, {Alexander F.} and Spivak, {Charles E.} and Lupica, {Carl R.} and Underhill, {Suzanne M.} and Amara, {Susan G.} and Andrii Domanskyi and Anttila, {Jenni E.} and Mikko Airavaara and Hope, {Bruce T.} and Hamra, {F. Kent} and Richie, {Christopher T.} and Harvey, {Brandon K.}",
note = "Funding Information: This study was supported by the Intramural Programs of the National Institute on Drug Abuse and National Institute of Mental Health . M.A., A.D., and J.E.A. were supported by the Academy of Finland . M.A. and A.D. were supported by 3iRegeneration Tekes . J.E.A. was supported by the University of Helsinki Doctoral Program in Drug Research . Transgenic LSL-Cas9 nickase rat production was supported by the UT Southwestern Mutant Rat Resource in Dallas. The NIDA Histology Core and Electrophysiology Core (EVEC) provided technical support. Dr. Francois Vautier oversaw the breeding of the transgenic rats. Doug Howard produced the AAV vectors. Reinis Svarcbahs assisted with the molecular biology in generating vectors. Dr. Deon Harvey made comments on the manuscript. Coronal brain schematics in figures are based on Swanson (2018) . Publisher Copyright: {\textcopyright} 2019",
year = "2019",
month = apr,
day = "3",
doi = "10.1016/j.neuron.2019.01.035",
language = "English (US)",
volume = "102",
pages = "105--119.e8",
journal = "Neuron",
issn = "0896-6273",
publisher = "Cell Press",
number = "1",
}