Aav5 ef1a dio eyfp

Manufactured by Addgene

AAV5-EF1a-DIO-EYFP is an adeno-associated virus (AAV) vector that expresses the enhanced yellow fluorescent protein (EYFP) gene under the control of the EF1α promoter. The DIO (double-floxed inverted open reading frame) configuration allows for Cre-dependent expression of the EYFP reporter.

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Lab products found in correlation

Aav5 ef1a dio hchr2 h134r eyfp, by Addgene (3 mentions) Aav5 hsyn dio hm4d gi mcherry, by Addgene (2 mentions) Aav5 hsyn dio hm3d gq mcherry, by Addgene (2 mentions) Aav dj ef1adiogcamp6m, by Addgene (2 mentions) Aav5 hsyn dio mcherry, by Addgene (1 mentions) Aav5 ef1a dio enphr 3.0 eyfp, by Addgene (1 mentions) Quintessential stereotaxic injector, by Stoelting (1 mentions) Stereotaxic frame, by Harvard Apparatus (1 mentions) Aav5 ef1a dio mcherry, by Addgene (1 mentions) Aav9 cag flex gfp, by Addgene (1 mentions)

6 protocols using aav5 ef1a dio eyfp

Viral Vector Toolbox Protocols

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The following AAV vectors (with a titer of >10¹²) were purchased from the vector core at the University of North Carolina at Chapel Hill (UNC Vector Core): AAV5-EF1a-DIO-hChR2(H134R)-EYFP, AAV5-EF1a-DIO-EYFP, and AAV-DJ-EF1aDIOGCaMP6m. The following AAV vectors were from Addgene: AAV5-hSyn-DIO-hM3D(Gq)-mCherry (no. 44361), AAV5-hSyn-DIO-hM4D(Gi)-mCherry (no. 44362), and AAV5-hSyn-DIO-mCherry (no. 50459).

Zhao Z.D., Han X., Chen R., Liu Y., Bhattacherjee A., Chen W, & Zhang Y. (2022). A molecularly defined D1 medium spiny neuron subtype negatively regulates cocaine addiction. Science Advances, 8(31), eabn3552.

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Cre-dependent Optogenetic Inhibition

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A Cre-dependent inhibitory optogenetic construct halorhodopsin (eNpHR, AAV5-Ef1a-DIO eNpHR 3.0-EYFP at titer ≥ 1×10¹³ vg/mL, Addgene plasmid # 26966) or an empty vector (control, AAV5- Ef1a-DIO EYFP at titer ≥ 1×10¹³ vg/mL, Addgene plasmid # 27056) were utilized. Both plasmids were obtained from Addgene as gifts from Dr. Karl Deisseroth.

Iglesias A.G., Chiu A.S., Wong J., Campus P., Li F., Liu Z.(., Patel S.A., Deisseroth K., Akil H., Burgess C.R, & Flagel S.B. (2023). Inhibition of dopamine neurons prevents incentive value encoding of a reward cue: With revelations from deep phenotyping. bioRxiv.

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Targeted AAV Transduction of Mouse Striatum

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SOM- cre mice were anesthetized with isoflurane and placed in a stereotaxic frame (Harvard Apparatus, Holliston, MA). A Quintessential Stereotaxic Injector (Stoelting, Wood Dale, IL) was used to inject 0.5 µl of AAV5-EF1a-DIO-EYFP (#27056, Addgene) into the striatum (+0.5 AP, 2 ML, 2.5 DV) at a speed of 0.1 µl/min. The pipette was held in place for at least 5 min after the injection. Following the surgery, the mice were given analgesics (buprenorphine, 0.08 mg/kg, i.p.).

Matityahu L., Malgady J.M., Schirelman M., Johansson Y., Wilking J.A., Silberberg G., Goldberg J.A, & Plotkin J.L. (2022). A tonic nicotinic brake controls spike timing in striatal spiny projection neurons. eLife, 11, e75829.

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Viral Transduction for Neurobehavioral Studies

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For immunostaining of LC fibers in the CB, 230 nl of AAV5-synP-DIO-eGFP-WPRE-hGH (Addgene,1*10^13 vg/ml) was injected into the LC of TH-Cre mice. For fiber photometry experiments targeting the CB vermis, 230 nl of AAV9-hSyn-GRAB_NE1m (Addgene,1 × 10¹³ vg/ml) was unilaterally injected in C57BL/6J mice. For optogenetic and chemogenetic experiments, 230 nl of AAV5-EF1a-DIO-eArch3.0-eYFP (Addgene,1 × 10¹³ vg/ml) and AAV5-hSyn-DIO-hM4D(Gi)-mCherry (Addgene,1 × 10¹³ vg/ml) were injected into the LC of TH-Cre mice, respectively; AAV5-EF1a-DIO-eYFP (230 nl, Addgene,1 × 10¹³ vg/ml) was injected into the LC of TH-cre mice used as controls.

Stanley A.T., Post M.R., Lacefield C., Sulzer D, & Miniaci M.C. (2023). Norepinephrine release in the cerebellum contributes to aversive learning. Nature Communications, 14, 4852.

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Viral Tracing and Manipulation in Neuroscience

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All viral vectors used in these studies have been extensively used in neuroscience. For anterograde tracing, AAV9-CAG-FLEX-GFP (Addgene, 51502) and AAV5-DIO-Synaptophysin-venus-GFP (Addgene, generated from plasmid 137188) were used. For monosynaptic rabies input circuit tracing, AAV5-Ef1A-DIO-GTB (Addgene, 27056), AAV5-Ef1A-DIO-TVA-mCherry (Addgene, generated from plasmid 37084) and EnvA-SAD-Rb-ΔG-GFP (32635, Salk Institute) were used. For chemogenetic studies, AAV5-EF1a-DIO-hM3Dq-mCherry (Addgene, 44631), AAV5-EF1a-DIO-hM4Di-mCherry (Addgene, 44632) or AAV5-EF1a-DIO-mCherry (Addgene, generated from plasmid 50462) were used. For optogenetic activation studies, AAV5-EF1a-DIO-hChR2(H134R)-EYFP (Addgene, 20298) or AAV5-EF1a-DIO-EYFP (Addgene, 27056) were used.

Schneeberger M., Brice N.L., Pellegrino K., Parolari L., Shaked J.T., Page K.J., Marchildon F., Barrows D.W., Carroll T.S., Topilko T., Mulligan V.M., Newman R., Doyle K., Bürli R., Barker D.F., Glen A., Ortuño M.J., Nectow A.R., Renier N., Cohen P., Carlton M., Heintz N, & Friedman J.M. (2022). Pharmacological targeting of glutamatergic neurons within the brainstem for weight reduction. Nature Metabolism, 4(11), 1495-1513.

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Diverse Viral Vectors for Optogenetics

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The following AAV vectors (with a titer of >10¹²) were purchased from UNC Vector Core: AAV5-EF1a-DIO-hChR2(H134R)-EYFP, AAV5-EF1a-DIO-EYFP, AAV-DJ-EF1a-DIO-GCaMP6m. The following AAV vectors were purchased from Addgene: AAV5-hSyn-DIO-hM3D(Gq)-mCherry (#44361), AAV5-hSyn-DIO-hM4D(Gi)-mCherry (#44362), AAV5-hSyn-DIO-mCherry (#50459), pAAV-flex-taCasp3-TEVp (#45580), pAAV-Ef1α-DIO eNpHR 3.0-EYFP (#26966), EnvA G-deleted Rabies-EGFP (Salk Institute).

Liu Y., Zhao Z.D., Xie G., Chen R, & Zhang Y. (2023). A molecularly distinct accumbal-to-lateral hypothalamic circuit modulates food seeking and consumption. bioRxiv.

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