Small-Scale Recombinant Adeno-Associated Virus Purification
Optiprep 5 M NaCl 5×TD Water Final volume 15 % 45 ml 36 ml 36 ml 63 ml 180 ml 30 % 60 ml – 24 ml 36 ml…
Expression of Multiple Functional RNAs or Proteins from One Viral Vector
Strategy Products Stoichiometry Advantages Disadvantages Potential applications References Fusion protein (Fig. 1a) Protein 1:1 predictable + Intracellular co-localization of the two proteins is guaranteed + Very space-efficient thus good for…
Controlling AAV Tropism in the Nervous System with Natural and Engineered Capsids
© Springer Science+Business Media New York 2016Fredric P. Manfredsson (ed.)Gene Therapy for Neurological DisordersMethods in Molecular Biology138210.1007/978-1-4939-3271-9_10 10. Controlling AAV Tropism in the Nervous System with Natural and Engineered Capsids Michael J. Castle1, 4, Heikki T. Turunen5, Luk H. Vandenberghe5…
Regulated Gene Therapy
Fig. 1 Schematic representation of the Tet-Off (left ) and Tet-On (right ) system, in the presence (higher panel ) or absence (lower panel ) of tetracycline (red squares). Tet-Off: the transactivator (tTA) is…
Directed Evolution of Adenoviruses
Fig. 1 The viral genomic sequence immediately surrounding the F421Y mutation (bold) is indicated. Note that the pol gene is on the complementary strand (translated) as the adenovirus genome is…
Introduction to Viral Vectors and Other Delivery Methods for Gene Therapy of the Nervous System
Vector Capacity Flexibility Considerations rAAV 4.7-5 kb Tropism can be changed by pseudotyping and directed evolution. The capsid may illicit an immune response when delivered peripherally, or with readministration centrally. …
Lentivirus Production and Purification
Fig. 1 GFP expression following transduction with a VSV-G pseudotyped LV vector. LV vectors produced using the current protocol efficiently transduces cells in vitro (a–c) and in vivo (d). SH-SY5Y…
Adeno-Associated Viral Gene Therapy for Retinal Disorders
Fig. 1 Basic vertebrate retinal structure image showing the various layers of the vertebrate retina with light passing through the ganglion cell layer (GCL), inner nuclear layer (INL) to the…
AAV Gene Therapy Strategies for Lysosomal Storage Disorders with Central Nervous System Involvement
Fig. 1 Lysosomal enzymes involved in the catabolism of GM1 ganglioside to GM3 ganglioside and the lysosomal storage disorders associated with loss of each enzyme Adeno-associated virus is a replication-deficient…
Gene Therapy in Spinal Muscular Atrophy (SMA) Models Using Intracerebroventricular Injection into Neonatal Mice
Fig. 1 Schematic of the exon 7 regulation within SMN pre-mRNA and trans-splicing strategy to replace the SMN2 exon 7. (a) SMN pre-mRNA is regulated by cis elements (within intron…
