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

Neuron | 2019

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.

Pubmed ID: 30792150 RIS Download

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This is a list of tools and resources that we have found mentioned in this publication.


Anti-Tyrosine Hydroxylase Antibody (antibody)

RRID:AB_390204

This polyclonal targets Tyrosine Hydroxylase

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β-Actin (8H10D10) Mouse mAb (antibody)

RRID:AB_2242334

This monoclonal targets beta-actin

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Anti-Tyrosine Hydroxylase Antibody, clone LNC1 (antibody)

RRID:AB_2201528

This monoclonal targets Tyrosine Hydroxylase

View all literature mentions

Purified anti-Cre Recombinase (antibody)

RRID:AB_2565079

This unknown targets Cre Recombinase

View all literature mentions

LE-Tg(Slc6a3-icre)6Ottc (organism)

RRID:RGD_9588578

Rattus norvegicus with name LE-Tg(Slc6a3-icre)6Ottc from RGD.

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Fiji (software resource)

RRID:SCR_002285

Software package as distribution of ImageJ and ImageJ2 together with Java, Java3D and plugins organized into coherent menu structure. Used to assist research in life sciences.

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PC12 (cell line)

RRID:CVCL_0481

Cell line PC12 is a Cancer cell line with a species of origin Rattus norvegicus

View all literature mentions

Anti-Tyrosine Hydroxylase Antibody, clone LNC1 (antibody)

RRID:AB_2201528

This monoclonal targets Tyrosine Hydroxylase

View all literature mentions

β-Actin (8H10D10) Mouse mAb (antibody)

RRID:AB_2242334

This monoclonal targets beta-actin

View all literature mentions

Anti-Tyrosine Hydroxylase Antibody (antibody)

RRID:AB_390204

This polyclonal targets Tyrosine Hydroxylase

View all literature mentions

Purified anti-Cre Recombinase (antibody)

RRID:AB_2565079

This unknown targets Cre Recombinase

View all literature mentions

Anti-Tyrosine Hydroxylase Antibody, clone LNC1 (antibody)

RRID:AB_2201528

This monoclonal targets Tyrosine Hydroxylase

View all literature mentions

LE.Cg-(ROSA26) em1(CAG-Cas9*D10A)Ottc (organism)

RRID:RRRC_00834

Rattus norvegicus with name LE.Cg-(ROSA26) em1(CAG-Cas9*D10A)Ottc from RGD.

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LE-Tg(DIO-mCherry)2Ottc (organism)

RRID:RRRC_00687

Rattus norvegicus with name LE-Tg(DIO-mCherry)2Ottc from RGD.

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LE-Tg(Slc6a3-icre)1Ottc (organism)

RRID:RRRC_00730

Rattus norvegicus with name LE-Tg(Slc6a3-icre)1Ottc from RGD.

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