東華大學圖書館 |

語系: 繁體中文

說明(常見問題)

回圖書館首頁

手機版館藏查詢

登入

回首頁

切換: 標籤 | MARC模式 | ISBD

In Vitro-Reconstituted Virus-Like-Pa...

Biddlecome, Adam Min.

FindBook

Google Book

Amazon

博客來

In Vitro-Reconstituted Virus-Like-Particle Delivery of Self-Amplifying RNA Genes.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	In Vitro-Reconstituted Virus-Like-Particle Delivery of Self-Amplifying RNA Genes./
作者:	Biddlecome, Adam Min.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2019,
面頁冊數:	87 p.
附註:	Source: Dissertations Abstracts International, Volume: 81-04, Section: B.
Contained By:	Dissertations Abstracts International81-04B.
標題:	Biochemistry. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=13899671
ISBN:	9781085677844

In Vitro-Reconstituted Virus-Like-Particle Delivery of Self-Amplifying RNA Genes.
Biddlecome, Adam Min.

In Vitro-Reconstituted Virus-Like-Particle Delivery of Self-Amplifying RNA Genes. - Ann Arbor : ProQuest Dissertations & Theses, 2019 - 87 p.

Source: Dissertations Abstracts International, Volume: 81-04, Section: B.

Thesis (Ph.D.)--University of California, Los Angeles, 2019.

This item must not be sold to any third party vendors.

The delivery of RNA genes has great potential in a range of therapeutic applications. A couple of main limitations for RNA-based therapies include its vulnerability to ribonuclease (RNase) digestion, and its transient and low expression levels due to its lack of amplification. Accordingly, a gene delivery platform that includes self-amplifying mRNA inside a protective capsid allowing for cell targeting and uptake could represent a major step forward in mRNA-based gene therapy.The self-amplifying nature of viral-encoded RNA-dependent RNA polymerases (RdRp) of Nodamura virus holds promise in the development of a platform for self-amplifying RNA gene delivery. To protect and assist in uptake of mRNA encoding an RdRp coupled to a gene of interest, plant virus-like-particles (VLPs) are a strong candidate. Cowpea chlorotic mottle virus (CCMV) capsid protein has been shown to be able to package in vitro heterologous mRNAs, and also to make the genes available to mammalian cells. To better characterize the uptake and amplification in mammalian cells of RdRp-associated genes (also called a replicon, because of its self-amplifying nature), both when naked or encapsidated, reporter genes such as enhanced yellow fluorescent protein (EYFP) or luciferase were used.In collaboration with the German pharmaceutical company Boehringer Ingelheim (B-I), evaluation of possible efficacy of this platform for a cancer vaccine was explored by incorporation of a model antigen (Ovalbumin epitope, SIINFEKL) sequence into the RNA1 of Nodamura virus. From in vitro studies involving incubation of OVA-replicon-VLPs, we observed that CCMV VLPs can be taken up by dendritic cells, where replication then occurs to promote increased production of antigen. Flow cytometry analysis revealed increased expression of maturation markers, and assaying the dendritic cell content by qPCR showed a significant number of SIINFEKL epitope-containing RNA. After these OVA-replicon-VLPs are injected into mice, a population of SIINFEKL-specific CD8+ (cytotoxic) T cells results.

ISBN: 9781085677844Subjects--Topical Terms:

518028
Biochemistry.
Subjects--Index Terms:

Dendritic cell activation

In Vitro-Reconstituted Virus-Like-Particle Delivery of Self-Amplifying RNA Genes.
LDR:03190nmm a2200349 4500 001 2272630
005 20201105110142.5
008 220629s2019 ||||||||||||||||| ||eng d
020 $a 9781085677844
035 $a (MiAaPQ)AAI13899671
035 $a AAI13899671
040 $a MiAaPQ $c MiAaPQ
100 1 $a Biddlecome, Adam Min. $3 3550056
245 1 0 $a In Vitro-Reconstituted Virus-Like-Particle Delivery of Self-Amplifying RNA Genes.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2019
300 $a 87 p.
500 $a Source: Dissertations Abstracts International, Volume: 81-04, Section: B.
500 $a Advisor: Gelbart, William M.
502 $a Thesis (Ph.D.)--University of California, Los Angeles, 2019.
506 $a This item must not be sold to any third party vendors.
520 $a The delivery of RNA genes has great potential in a range of therapeutic applications. A couple of main limitations for RNA-based therapies include its vulnerability to ribonuclease (RNase) digestion, and its transient and low expression levels due to its lack of amplification. Accordingly, a gene delivery platform that includes self-amplifying mRNA inside a protective capsid allowing for cell targeting and uptake could represent a major step forward in mRNA-based gene therapy.The self-amplifying nature of viral-encoded RNA-dependent RNA polymerases (RdRp) of Nodamura virus holds promise in the development of a platform for self-amplifying RNA gene delivery. To protect and assist in uptake of mRNA encoding an RdRp coupled to a gene of interest, plant virus-like-particles (VLPs) are a strong candidate. Cowpea chlorotic mottle virus (CCMV) capsid protein has been shown to be able to package in vitro heterologous mRNAs, and also to make the genes available to mammalian cells. To better characterize the uptake and amplification in mammalian cells of RdRp-associated genes (also called a replicon, because of its self-amplifying nature), both when naked or encapsidated, reporter genes such as enhanced yellow fluorescent protein (EYFP) or luciferase were used.In collaboration with the German pharmaceutical company Boehringer Ingelheim (B-I), evaluation of possible efficacy of this platform for a cancer vaccine was explored by incorporation of a model antigen (Ovalbumin epitope, SIINFEKL) sequence into the RNA1 of Nodamura virus. From in vitro studies involving incubation of OVA-replicon-VLPs, we observed that CCMV VLPs can be taken up by dendritic cells, where replication then occurs to promote increased production of antigen. Flow cytometry analysis revealed increased expression of maturation markers, and assaying the dendritic cell content by qPCR showed a significant number of SIINFEKL epitope-containing RNA. After these OVA-replicon-VLPs are injected into mice, a population of SIINFEKL-specific CD8+ (cytotoxic) T cells results.
590 $a School code: 0031.
650 4 $a Biochemistry. $3 518028
650 4 $a Virology. $3 642304
650 4 $a Genetics. $3 530508
653 $a Dendritic cell activation
653 $a Self-replicating
653 $a Virus-like particles
690 $a 0487
690 $a 0720
690 $a 0369
710 2 $a University of California, Los Angeles. $b Biochemistry, Molecular and Structural Biology 0090. $3 3550057
773 0 $t Dissertations Abstracts International $g 81-04B.
790 $a 0031
791 $a Ph.D.
792 $a 2019
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=13899671