دورية أكاديمية
Cell-Specific Single Viral Vector CRISPR/Cas9 Editing and Genetically Encoded Tool Delivery in the Central and Peripheral Nervous Systems.
| العنوان: | Cell-Specific Single Viral Vector CRISPR/Cas9 Editing and Genetically Encoded Tool Delivery in the Central and Peripheral Nervous Systems. |
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| المؤلفون: | Moffa JC; Washington University Pain Center, Department of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri 63110.; Washington University Medical Scientist Training Program, Washington University School of Medicine, St. Louis, Missouri 63110., Bland IN; Washington University Pain Center, Department of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri 63110., Tooley JR; Washington University Pain Center, Department of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri 63110.; Washington University Division of Biological and Behavioral Sciences, Washington University School of Medicine, St. Louis, Missouri 63110., Kalyanaraman V; Washington University Pain Center, Department of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri 63110., Heitmeier M; Washington University Pain Center, Department of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri 63110., Creed MC; Washington University Pain Center, Department of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri 63110.; Departments of Neuroscience, Psychiatry, and Biomedical Engineering, Washington University School of Medicine, St. Louis, Missouri 63110., Copits BA; Washington University Pain Center, Department of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri 63110 bcopits@wustl.edu. |
| المصدر: | ENeuro [eNeuro] 2024 Jul 05; Vol. 11 (7). Date of Electronic Publication: 2024 Jul 05 (Print Publication: 2024). |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Society for Neuroscience Country of Publication: United States NLM ID: 101647362 Publication Model: Electronic-Print Cited Medium: Internet ISSN: 2373-2822 (Electronic) Linking ISSN: 23732822 NLM ISO Abbreviation: eNeuro Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: [Washington, DC] : Society for Neuroscience, [2014]- |
| مواضيع طبية MeSH: | CRISPR-Cas Systems* , Dependovirus*/genetics , Gene Editing*/methods , Genetic Vectors*, Animals ; Mice ; Optogenetics/methods ; Central Nervous System/metabolism ; Peripheral Nervous System/metabolism ; Male ; Mice, Inbred C57BL ; Neurons/metabolism ; Female ; Mice, Transgenic |
| مستخلص: | CRISPR/Cas9 gene editing represents an exciting avenue to study genes of unknown function and can be combined with genetically encoded tools such as fluorescent proteins, channelrhodopsins, DREADDs, and various biosensors to more deeply probe the function of these genes in different cell types. However, current strategies to also manipulate or visualize edited cells are challenging due to the large size of Cas9 proteins and the limited packaging capacity of adeno-associated viruses (AAVs). To overcome these constraints, we developed an alternative gene editing strategy using a single AAV vector and mouse lines that express Cre-dependent Cas9 to achieve efficient cell-type specific editing across the nervous system. Expressing Cre-dependent Cas9 from a genomic locus affords space to package guide RNAs for gene editing together with Cre-dependent, genetically encoded tools to manipulate, map, or monitor neurons using a single virus. We validated this strategy with three common tools in neuroscience: ChRonos, a channelrhodopsin, for studying synaptic transmission using optogenetics, GCaMP8f for recording Ca 2+ transients using photometry, and mCherry for tracing axonal projections. We tested these tools in multiple brain regions and cell types, including GABAergic neurons in the nucleus accumbens, glutamatergic neurons projecting from the ventral pallidum to the lateral habenula, dopaminergic neurons in the ventral tegmental area, and proprioceptive neurons in the periphery. This flexible approach could help identify and test the function of novel genes affecting synaptic transmission, circuit activity, or morphology with a single viral injection. Competing Interests: The authors declare no competing financial interests. (Copyright © 2024 Moffa et al.) |
| التعليقات: | Update of: bioRxiv. 2023 Oct 10:2023.10.10.561249. doi: 10.1101/2023.10.10.561249. (PMID: 37873336) |
| References: | Methods Mol Biol. 2014;1183:221-42. (PMID: 25023312) Nat Genet. 2016 Oct;48(10):1112-8. (PMID: 27618451) Nat Commun. 2022 Dec 2;13(1):7433. (PMID: 36460649) Lab Anim (NY). 2011 May;40(5):155-60. (PMID: 21508954) Neuroscience. 2014 Dec 12;282:109-21. (PMID: 25073045) Front Neuroinform. 2018 Apr 04;12:14. (PMID: 29670519) Nat Commun. 2019 Sep 6;10(1):4056. (PMID: 31492834) Hum Gene Ther. 1996 Nov 10;7(17):2101-12. (PMID: 8934224) PLoS Biol. 2020 Apr 10;18(4):e3000665. (PMID: 32275651) Nature. 2023 Jul;619(7969):332-337. (PMID: 37380765) Dev Dyn. 2002 Jun;224(2):135-43. (PMID: 12112467) Neuron. 2023 Jun 7;111(11):1776-1794.e10. (PMID: 37028432) Trends Biochem Sci. 1998 Jun;23(6):198-9. (PMID: 9644970) Transl Psychiatry. 2017 Jan 24;7(1):e1013. (PMID: 28117842) Science. 2014 Jan 3;343(6166):84-87. (PMID: 24336571) Nat Genet. 2019 Nov;51(11):1645-1651. (PMID: 31659324) Curr Biol. 2021 Oct 11;31(19):4388-4396.e5. (PMID: 34388372) Elife. 2021 Mar 29;10:. (PMID: 33779547) Hum Gene Ther. 2012 Aug;23(8):847-58. (PMID: 22545762) Nat Biotechnol. 2016 Feb;34(2):184-191. (PMID: 26780180) Science. 2008 Sep 19;321(5896):1690-2. (PMID: 18802002) Elife. 2022 Apr 14;11:. (PMID: 35420982) Nature. 2022 Aug;608(7923):586-592. (PMID: 35859170) Nat Biotechnol. 2015 Jan;33(1):102-6. (PMID: 25326897) Nat Commun. 2018 Feb 27;9(1):849. (PMID: 29487284) Proc Natl Acad Sci U S A. 2009 May 5;106(18):7281-8. (PMID: 19342487) Nat Methods. 2014 Mar;11(3):338-46. (PMID: 24509633) Neuron. 2011 Jul 14;71(1):142-54. (PMID: 21745644) Nature. 2023 Mar;615(7954):884-891. (PMID: 36922596) Nature. 2021 Oct;598(7882):646-651. (PMID: 34646022) Mol Psychiatry. 2022 Dec;27(12):4994-5006. (PMID: 36100669) Nature. 2006 Aug 24;442(7105):929-33. (PMID: 16906136) Annu Rev Biophys. 2017 May 22;46:505-529. (PMID: 28375731) Addict Biol. 2023 Jan;28(1):e13247. (PMID: 36577719) Hear Res. 2020 Sep 1;394:107912. (PMID: 32067799) Brain Res Brain Res Rev. 1997 Dec;25(3):312-34. (PMID: 9495561) Neuropsychopharmacology. 2020 Nov;45(12):2020-2029. (PMID: 32585679) Nat Neurosci. 2017 Aug;20(8):1172-1179. (PMID: 28671695) Mol Brain. 2022 Mar 4;15(1):21. (PMID: 35246205) Nucleic Acids Res. 2018 Jul 2;46(W1):W242-W245. (PMID: 29762716) Neuron. 2019 Apr 03;102(1):105-119.e8. (PMID: 30792150) Sci Adv. 2023 Aug 11;9(32):eadg8869. (PMID: 37566654) Genes Dev. 2006 Feb 1;20(3):355-67. (PMID: 16452507) Cell Rep. 2016 Nov 22;17(9):2163-2172. (PMID: 27880894) J Virol. 2005 Aug;79(15):9933-44. (PMID: 16014954) Mamm Genome. 2000 Mar;11(3):196-205. (PMID: 10723724) Nat Protoc. 2017 Apr;12(4):828-863. (PMID: 28333914) Proc Natl Acad Sci U S A. 2010 Jul 27;107(30):13491-6. (PMID: 20616081) J Biol Chem. 2014 Aug 1;289(31):21312-24. (PMID: 24907273) Nat Methods. 2021 Jan;18(1):100-106. (PMID: 33318659) Elife. 2021 Mar 10;10:. (PMID: 33689678) Sci Rep. 2021 Dec 20;11(1):24212. (PMID: 34930955) Neuron. 2023 Aug 2;111(15):2282-2311. (PMID: 37201524) EMBO J. 2001 Sep 3;20(17):4987-97. (PMID: 11532962) Cerebrovasc Dis. 2009;27 Suppl 1:162-7. (PMID: 19342847) Dev Biol. 2000 Jun 15;222(2):296-306. (PMID: 10837119) Sci Rep. 2019 Mar 12;9(1):4194. (PMID: 30862905) Cell Rep. 2022 Jan 04;38(1):110196. (PMID: 34986352) Neuron. 2023 Oct 4;111(19):3053-3067.e10. (PMID: 37480845) Cell. 2014 Oct 9;159(2):440-55. (PMID: 25263330) Nature. 2012 Oct 11;490(7419):262-6. (PMID: 23034651) Neuron. 2009 Jun 25;62(6):826-38. (PMID: 19555651) J Neurosci. 2010 Jun 16;30(24):8229-33. (PMID: 20554874) J Neurochem. 2011 Nov;119(3):544-54. (PMID: 21883221) Science. 2012 Aug 17;337(6096):816-21. (PMID: 22745249) Biol Psychiatry. 2018 Jun 15;83(12):1012-1023. (PMID: 29452828) Neuron. 2017 May 3;94(3):611-625.e4. (PMID: 28472659) Genesis. 2004 Oct;40(2):67-73. (PMID: 15452869) Elife. 2021 Jun 08;10:. (PMID: 34100715) Neuron. 2021 Jun 2;109(11):1791-1809.e11. (PMID: 33979635) Nat Neurosci. 2008 Jun;11(6):721-8. (PMID: 18454144) Cell Rep. 2020 Mar 24;30(12):4303-4316.e6. (PMID: 32209486) PLoS Biol. 2005 May;3(5):e159. (PMID: 15836427) Gene Ther. 2023 May;30(5):463-468. (PMID: 34176926) Neuron. 2019 Aug 21;103(4):583-597.e8. (PMID: 31272828) Genes Dev. 2015 Jul 15;29(14):1576-85. (PMID: 26178787) Nat Protoc. 2019 Feb;14(2):379-414. (PMID: 30626963) |
| معلومات مُعتمدة: | R01 DA058755 United States DA NIDA NIH HHS; R01 NS130046 United States NS NINDS NIH HHS |
| فهرسة مساهمة: | Keywords: CRISPR/Cas9; gene editing; imaging; optogenetics; photometry; tool |
| تواريخ الأحداث: | Date Created: 20240613 Date Completed: 20240705 Latest Revision: 20240710 |
| رمز التحديث: | 20240710 |
| مُعرف محوري في PubMed: | PMC11228695 |
| DOI: | 10.1523/ENEURO.0438-23.2024 |
| PMID: | 38871457 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2373-2822 |
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| DOI: | 10.1523/ENEURO.0438-23.2024 |