Engineered newcastle disease virus vector and uses thereof
AssigneeUniversity of Guelph
InventorsSarah Wootton, Leonardo Susta, Byram Bridle, Pierre Major, Lisa Santry, Yanlong Pei
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Abstract
An engineered Newcastle Disease Virus (NDV) vector is provided. In particular, the present disclosure provides methods of treating or preventing a disease such as cancer, or an infectious disease, or methods for eliciting an immune response, with the engineered NDV vector. The engineered NDV vector provided herein is useful as an immunogenic composition, an oncolytic agent, or a vaccine.
Claims19
An engineered Newcastle Disease Virus (NDV) vector comprising a nucleic acid having a nucleic acid sequence that is at least 95% or 100% identical to the nucleic acid sequence of any one of SEQ ID NO:
- 1-5, 9, 10, 18, 19, 23, 27, or 42, wherein the nucleic acid comprises or further comprises at least one heterologous nucleic acid segment encoding a therapeutic agent operably linked to a promoter capable of expressing the segment in a host cell, wherein the nucleic acid comprises a nucleic acid sequence encoding an L protein having a stabilizing segment, and wherein the nucleic acid comprises XbaI and MluI restriction endonuclease sites between nucleic acid sequence encoding phosphoprotein and matrix protein.
An engineered Newcastle Disease Virus (NDV) vector comprising a nucleic acid having a nucleic acid sequence encoding an L protein having a stabilizing segment, a chimeric F protein, and a chimeric HN protein, wherein the chimeric F protein comprises avian paramyxovirus 5 (APMV5) F protein segment thereof at the N-terminus and an NDV F protein segment at the C-terminus, and wherein the chimeric HN protein comprises an NDV HN protein segment at the N-terminus and an AMPV5 HN protein segment at the C-terminus.
A method for selecting an engineered NDV vector genome comprising a stabilizing segment in L gene, the method comprises:
- a) growing bacterial cells comprising an engineered NDV vector genome in a growth medium broth;
- b) growing the bacterial cells on an agar-growth medium, wherein the agar-growth medium comprises a selection agent;
- c) identifying small bacterial cell colonies having about 0.5 mm to about 1 mm in diameter after at least 24 hours of growth;
- d) repeating step a) to step c) two to nine times to enrich for small bacterial cell colonies;
- and
- e) isolating the engineered NDV vector genome from the small bacterial cells colonies,
- wherein the small bacterial cells colonies comprise stable engineered NDV vector genome having the stabilizing segment in L gene.
Dependencies are inferred from the claim text.