Synthetic combinatorial aav capsid library for targeted gene therapy

Inactive Publication Date: 2016-12-22
UNIV OF FLORIDA RES FOUNDATION INC
View PDF1 Cites 55 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0022]In various gene therapy applications of the disclosed invention, the nucleic acid contained within the AAV vector will preferably further include a DNA sequence that encodes one or more diagnostic, therapeutic, and/or prophylactic molecules. In such embodiments, the nucleic acid preferably further includes one or m

Problems solved by technology

In addition, gene synthesis allows virtual recombination between naturally-occurring sequences at

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Synthetic combinatorial aav capsid library for targeted gene therapy
  • Synthetic combinatorial aav capsid library for targeted gene therapy
  • Synthetic combinatorial aav capsid library for targeted gene therapy

Examples

Experimental program
Comparison scheme
Effect test

example 1

AAV Vector Library Construction

[0149]Methodologies to improve existing AAV vectors for gene therapy include either rational approaches or directed evolution to derive capsid variants characterized by superior transduction efficiencies in targeted tissues. In the present invention, both approaches were integrated in one unified design strategy of “virtual family shuffling” to derive a combinatorial capsid library whereby only variable regions on the surface of the capsid are modified. Individual sub-libraries were first assembled in order to pre-select compatible amino acid residues within restricted surface-exposed regions to minimize the generation of dead-end variants. Subsequently, the successful families were interbred to derive a combined library of about 1×108 complexity. Next-Gen sequencing of the packaged viral DNA revealed capsid surface areas susceptible to directed evolution thus providing guidance for future designs. The utility of the library in gene therapy application...

example 2

Integrating Combinatorial and Rational Approaches to Derive Novel Aav Variants

[0154]AAV is a single-stranded DNA virus belonging to the Parvoviridae family (Muzyczka and Berns, 2001). AAV-derived vectors are promising tools for human gene therapy applications because of their absence of pathogenicity, episomal localization and stable transgene expression (Wu et al., 2006). However, significant limitations to the clinical use of AAV are its promiscuity and its susceptibility to neutralization by human antibodies (Louis-Jeune et al., 2013). Both of these limitations are determined by the nature of amino acid residues exposed at the surface of the capsid. Two main strategies are generally employed to improve AAV vectors: 1) mutagenizing capsid residues to facilitate binding, entry, and / or intracellular trafficking through a rational approach based on the knowledge of virus biology (Wu et al., 2000; Zhong et al., 2008; Lochrie et al., 2005; Aslanidi et al., 2012; Li et al., 2012; Gabrie...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

Disclosed are compositions and methods for producing modified adeno-associated virus (AAV) cap genes and combinatorial libraries of chimeric AAV vectors and virions; selecting for virions displaying cell-specific tropisms; and, in certain embodiments, producing helper vectors containing one or more modified AAV cap genes. The synthetic combinatorial AAV capsid libraries of the invention are useful in introducing into selected target host cells one or more nucleic acid molecules. The viral vectors and genetic constructs disclosed herein are also useful in a variety of diagnostic and/or therapeutic gene-therapy regimens.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]The present application claims priority to U.S. Provisional Patent Appl. No. 61 / 883,063, filed Sep. 26, 2013 (pending; Atty. Dkt. No. 36689.341). The content of which is specifically incorporated herein in its entirety by express reference thereto.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]This invention was made with government support under Grant Nos. HL-097088 and GM-082946 awarded by the National Institutes of Health. The government has certain rights in the invention.NAMES OF THE PARTIES TO A JOINT RESEARCH AGREEMENT[0003]Not Applicable.BACKGROUND OF THE INVENTION[0004]Field of the Invention[0005]The present invention relates to the fields of molecular biology, medicine, and virology. More particularly, the invention relates to compositions and methods for producing modified adeno-associated virus (AAV) cap genes and combinatorial libraries of chimeric AAV vectors and virions; and in particular embodiments, ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/86C12N7/00C07K14/005
CPCC12N15/86C07K14/005C12N2750/14143C12N2750/14122A61K48/00C12N7/00C12N2750/14145
Inventor MARSIC, DAMIENZOLOTUKHIN, SERGEIAGBANDJE-MCKENNA, MAVIS
Owner UNIV OF FLORIDA RES FOUNDATION INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap