title: A Simple Alternative to Stereotactic Injection for Brain Specific Knockdown of miRNA.
abstract: MicroRNAs (miRNAs) are key regulators of gene expression. In the brain, vital processes like neurodevelopment and neuronal functions depend on the correct expression of microRNAs. Perturbation of microRNAs in the brain can be used to model neurodegenerative diseases by modulating neuronal cell death. Currently, stereotactic injection is used to deliver miRNA knockdown agents to specific location in the brain. Here, we discuss strategies to design antagomirs against miRNA with locked nucleotide modifications (LNA). Subsequently describe a method for brain specific delivery of antagomirs, uniformly across different regions of the brain. This method is simple and widely applicable since it overcomes the surgery, associated injury and limitation of local delivery in stereotactic injections. We prepared a complex of neurotropic, cell-penetrating peptide Rabies Virus Glycoprotein (RVG) with antagomir against miRNA-29 and injected through tail vein, to specifically deliver in the brain. The antagomir design incorporated features that allow specific targeting of the miRNA and formation of non-covalent complexes with the peptide. The knock-down of the miRNA in neuronal cells, resulted in apoptotic cell death and associated behavioural defects. Thus, the method can be used for acute models of neuro-degeneration through the perturbation of miRNAs.
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, New Delhi, India.
authors.forename: Hemant
authors.initials: H
authors.lastname: Suryawanshi
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, New Delhi, India; Academy of Scientific and Innovative Research (AcSIR), New Delhi, India.
authors.forename: Mayuresh Anant
authors.initials: MA
authors.lastname: Sarangdhar
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, New Delhi, India; Academy of Scientific and Innovative Research (AcSIR), New Delhi, India.
authors.forename: Manika
authors.initials: M
authors.lastname: Vij
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, New Delhi, India.
authors.forename: Reema
authors.initials: R
authors.lastname: Roshan
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, New Delhi, India.
authors.forename: Vijay Pal
authors.initials: VP
authors.lastname: Singh
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, New Delhi, India.
authors.forename: Munia
authors.initials: M
authors.lastname: Ganguli
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, New Delhi, India; Academy of Scientific and Innovative Research (AcSIR), New Delhi, India; beena@igib.in.
authors.forename: Beena
authors.initials: B
authors.lastname: Pillai
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D055785:Gene Knockdown Techniques / Q000379:methods*; D006023:Glycoproteins / Q000008:administration & dosage / Q000235:genetics; D051379:Mice; D035683:MicroRNAs / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000235:genetics; D013238:Stereotaxic Techniques; D014162:Transfection / Q000379:methods; D014764:Viral Proteins / Q000008:administration & dosage / Q000235:genetics
chemical_list: D006023:Glycoproteins; C547014:MIRN29 microRNA, mouse; D035683:MicroRNAs; D016376:Oligonucleotides, Antisense; D014764:Viral Proteins