A Novel Fluorescent-Based Screen to Identify Small Synthetic Internal Ribosome Entry Site (IRES) Elements
- 技术优势
- Two-color reporter system to rapidly and efficiently determine hits Small synthetic IRES elements are changeable in length and sequence to accommodate a random mutagenesis approach or a directed design approach Small IRES size (<60 nt) allows for the incorporation of larger (or multiple) genes into a single retroviral vector for gene therapeutics Utilization of protoplast fusion for plasmid delivery imparts greater efficiency and dramatically lowers background noise
- 技术应用
- For the use in IRES element detection Identify trans-acting translation factors, alter cellular metabolism and augment current gene therapy applications
- 详细技术说明
- Researchers at UCLA have created an assay to identify IRES elements from a library of bicistronic plasmids encoding two fluorescent proteins separated by a small (<60 nt) synthetic intervening sequence of changeable length. By using fluorescence-activated cell-sorting techniques, it is possible to determine candidate IRES element sequences for further analysis.
- *Abstract
-
Researchers at UCLA have developed a novel, two-color fluorescent-based screen to identify small, synthetic internal ribosome entry site (IRES) elements in vivo. The system is also capable of quickly and efficiently identifying the IRES nature of any unknown natural or synthetic RNA element. Additionally, the synthetic small (~50 nt) IRES elements could be used for biotechnological purposes such as multiple protein expression from a single expression vector.
- *IP Issue Date
- Dec 21, 2004
- *Principal Investigation
-
Name: Asim Dasgupta
Department:
Name: Arun Venkatesan
Department:
- 附加资料
- Patent Number: US6833254B2
Application Number: US200287171A
Inventor: Dasgupta, Asim | Venkatesan, Arun
Priority Date: 1 Mar 2001
Priority Number: US6833254B2
Application Date: 1 Mar 2002
Publication Date: 21 Dec 2004
IPC Current: C12N001510 | C12N001521
US Class: 4350691 | 435005 | 435039 | 4353201 | 435334 | 435006
Assignee Applicant: The Regents of the University of California
Title: Method to identify IRES elements
Usefulness: Method to identify IRES elements
Summary: The method is useful for identifying trans-acting translation factors or IRES elements, which are useful for controlling viral infection in a cell or regulating cellular metabolism.
Novelty: Identifying an internal ribosome entry site (IRES) element for controlling viral infection or regulating metabolism in cells by employing a bicistronic system for reporter proteins involving both cap- and IRES-mediated translation
- 主要类别
- 生物医学
- 细分类别
- DNA /基因工程
- 申请号码
- 6833254
- 其他
-
State Of Development
The screen has been designed, tested and validated and two candidates, with no sequence or structural homology to known viral and cellular IRES elements, were shown to specifically block poliovirus and hepatitis C virus IRES-mediated translation.
Background
Internal ribosome entry site (IRES)-mediated translation initiates and assembles translation machinery at a site close to the start codon in a manner that does not require the traditional eukaryotic 5 nucleotide cap or eIF4E (cap-binding protein). Initially discovered in picornaviruses (poliovirus, encephalomyocarditis virus, rhinovirus, etc.), viral IRES-mediated translation relies on virus-specific combinations of transcription factors, trans-acting factors that help stabilize the IRES structure and the IRES elements themselves, which can span hundreds of nucleotides in length and have complicated secondary and tertiary structures. The complex way in which the IRES elements mediate translation is of great importance to researchers seeking insight into viral gene expression and ultimately, anti-viral therapeutics.
Related Materials
Tech ID/UC Case
20057/2001-202-0
Related Cases
2001-202-0
- 国家/地区
- 美国
