Data for: An AAV-CRISPR/Cas9-strategy for gene editing across divergent rodent species: Targeting neural oxytocin receptors as a proof of concept

A major issue in neuroscience is the poor translatability of research results from preclinical studies in animals to clinical outcomes. Comparative neuroscience can overcome this barrier by studying multiple species to differentiate between species-specific and general mechanisms of neural circuit functioning. Targeted manipulation of neural circuits often depends on genetic dissection, and use of this technique has been restricted to only a few model species, limiting its application in comparative research. However, ongoing advances in genomics make genetic dissection attainable in a growing number of species. To demonstrate the potential of comparative gene editing approaches, we developed a viral-mediated CRISPR/Cas9 strategy that is predicted to target the oxytocin receptor (Oxtr) gene in >80 rodent species. This strategy specifically reduced OXTR levels in all evaluated species (n=6), without causing gross neuronal toxicity. Thus, we show that CRISPR/Cas9-based tools can functi..., OXTR and AVPR1A autoradiography
 
Fresh frozen brains were sectioned on a cryostat (Epredia Cryostar NX-70, Thermo-Scientific, MA, USA) at 20 μm, mounted on Superfrost Plus slides (Fisher Scientific) and stored at -80C until use. Autoradiography was performed as previously described (39). Briefly, slides were thawed and fixed for two minutes in 0.1% paraformaldehyde in phospate-buffered saline (PBS) for two minutes, washed in 50 mM Tris in PBS (pH 7.4, 2x10 minutes) and incubated in 50 mM Tris buffer, supplemented with 0.1% BSA and 50 pM I125-OVTA (2200 Ci/mmol, ornithine vasotocin analog, #NEX254010UC, PerkinElmer, MA, USA) or 50 pM I125-AVP (2200 Ci/mmol, linear arginine vasopressin, #NEX310010UC, PerkinElmer) at room temperature for one hour. Unbound ligand was removed by washing in 50 mM Tris with 0.2% MgCl2 at 4 degrees (4x5 minutes) and 30 minutes at RT. Slides were dipped in miliQ, dried, and placed in a cassette with BioMax MR film (Sigma-Aldrich, MO, USA). After 7 days, films w..., Tables (.csv) can be opened with any standard software, such as excel. Code is written for R/R-studo