125 ovta  (Revvity)

Journal: Science Advances

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

doi: 10.1126/sciadv.adf4950

Figure Lengend Snippet: ( A ) Schematic depicting the two conserved regions in the Oxtr coding sequence in four rodent species that are amenable to CRISPR/Cas9–mediated mutagenesis. ( B ) Schematics illustrating the major elements within the AAV plasmids, the AAV particles and how they are infused in the brain, as well as how the CRISPR/Cas9 technique functions. ( C ) Image of agarose gel evidencing gene editing after infusion of AAV-ΔOXTR, but not AAV-CTRL. ( D ) Top images are representative I 125 -OVTA autoradiograms of brain sections of AAV-CRISPR/Cas9–injected prairie voles, and bottom images are adjacent brain sections that show viral-induced eGFP fluorescence. Black arrows indicate the target area. ( E ) Quantification of OXTR levels in AAV-ΔOXTR–injected hemispheres and AAV-CTRL–injected hemispheres. Paired t tests, ΔOXTR.1: N = 8, *** P = 0.0001; ΔOXTR.2: N = 8, *** P = 4.36 × 10 −5 . ( F ) Images of native eGFP fluorescence, immunofluorescence of NeuN protein, and 4′,6-diamidino-2-phenylindole (DAPI) staining in AAV-ΔOXTR–infected tissue on the left and quantification of Neun + cells in AAV- and sham-injected tissue. Paired t test: N = 8, P = 0.6. Scale bar, 20 μm. NS, nonsignificance. ( G ) Representative image of spCas9 mRNA expression in the prairie vole. Scale bar, 100 μm.

Article Snippet: Briefly, slides were thawed and fixed for 2 min in 0.1% paraformaldehyde in phosphate-buffered saline (PBS) for 2 min, washed in 50 mM tris in PBS (pH 7.4, 2 × 10 min), and incubated in 50 mM tris buffer, supplemented with 0.1% bovine serum albumin and 50 pM I 125 -OVTA (2200 Ci/mmol, ornithine vasotocin analog, #NEX254010UC, PerkinElmer, MA, USA) or 50 pM I 125 -AVP (2200 Ci/mmol, linear arginine vasopressin, #NEX310010UC, PerkinElmer) at room temperature (RT) for 1 hour.

Techniques: Sequencing, CRISPR, Mutagenesis, Agarose Gel Electrophoresis, Injection, Fluorescence, Immunofluorescence, Staining, Infection, Expressing

Journal: Science Advances

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

doi: 10.1126/sciadv.adf4950

Figure Lengend Snippet: Representative images of I 125 -OVTA autoradiograms of AAV-CRISPR/Cas9–injected ( A ) spiny mouse (AC), ( B ) golden hamster (MA), ( C ) house mouse (MM), ( D ) California deer mouse (PC), and ( E ) Norway rat (RN). ( F ) Quantification of OXTR levels in AAV-ΔOXTR–injected hemispheres and AAV-CTRL–injected hemispheres per species. Paired t tests: AC: N = 7, *** P = 4.57 × 10 −5 ; MA: N = 6, *** P = 0.0001; MM: N = 7, *** P = 4.1 × 10 −5 ; PC: N = 6, ** P = 0.006; RN: N = 10, *** P = 0.0001. Black arrows depict the target areas (NAc, nucleus accumbens; EC, endopiriform cortex; VMH, ventromedial hypothalamus; LS, lateral septum; CeA, central amygdala).

Article Snippet: Briefly, slides were thawed and fixed for 2 min in 0.1% paraformaldehyde in phosphate-buffered saline (PBS) for 2 min, washed in 50 mM tris in PBS (pH 7.4, 2 × 10 min), and incubated in 50 mM tris buffer, supplemented with 0.1% bovine serum albumin and 50 pM I 125 -OVTA (2200 Ci/mmol, ornithine vasotocin analog, #NEX254010UC, PerkinElmer, MA, USA) or 50 pM I 125 -AVP (2200 Ci/mmol, linear arginine vasopressin, #NEX310010UC, PerkinElmer) at room temperature (RT) for 1 hour.

Techniques: CRISPR, Injection

Journal: Neuron

Article Title: Wireless Optogenetic Stimulation of Oxytocin Neurons in a Semi-natural Setup Dynamically Elevates Both Pro-social and Agonistic Behaviors

doi: 10.1016/j.neuron.2020.05.028

Figure Lengend Snippet:

Article Snippet: Ornithine Vasotocin Analog (OVTA) , Perkin Elmer , NEX254050UC.

Techniques: Virus, Recombinant, Software, Amplification, Adhesive