Optimizing Expression of Recombinant Protein in Mammalian Cells

Use of an optimized “super core promoter” to enhance gene expression.The engineered core promoter is significantly stronger than known core promoters that are considered the strongest, such as AdML and CMV.Increases transcription/gene expression both in vitro and in in vivo transfections.

Instead of modifying upstream promoter and enhancer elements, UC San Diego researchers have developed an innovative technology based on combining elements of a eukaryotic core promoter to achieve significantly higher expression of recombinant proteins and reporters in mammalian cells. The inventors have designed, constructed, and optimized an extremely strong synthetic core promoter called the “super core promoter” (SCP). This technology could greatly enhance the expression of a wide variety of recombinant proteins and reporters in mammalian cells, in vitro and in vivo.

Patent Number: US7968698B2
Application Number: US2007920986A
Inventor: Kadonaga, James T. | Gershon, Tamar
Priority Date: 25 May 2005
Priority Number: US7968698B2
Application Date: 23 Nov 2007
Publication Date: 28 Jun 2011
IPC Current: C07H002104
US Class: 5360241 | 42409321 | 4350691 | 4353201
Assignee Applicant: The Regents of the University of California
Title: Optimized core promoters and uses thereof
Usefulness: Optimized core promoters and uses thereof
Summary: The construct, vector, promoter, and method are useful for increasing production of a protein in a metazoan cell.
Novelty: New core promoter construct comprising core promoter initiator element and core promoter motif ten element, useful for increasing production of a protein in a metazoan cell

化工/材料

化工/材料應用

7968698

State Of Development

SCP has been extensively characterized. Some of the salient features of SCP include the following:

1. SCP transcribes more efficiently than adenovirus major late (AdML) and the cytomegalovirus (CMV) core promoters.
2. In transient transfection and gene reporter assays in HeLa S3 and CHO cell lines, SCP is five to eight times stronger than AdML or CMV core promoters.
3. DNaseI footprinting experiments show that purified components of basal transcription machinery, such as TFIID, bind SCP with higher affinity than CMV or AdML core promoters.

Related Materials

• Lim CY, Santoso B, Boulay T, Dong E, Ohler U, Kadonaga JT. The MTE, A New Core Promoter Element for Transcription by RNA Polymerase II. Genes Dev. 2004; 18(13): p.1606-17.
• Juven-Gershon, T., S. Cheng, and Kadonaga JT. Rational Design of a Super Core Promoter that Enhances Gene Expression. Nat Methods, 2006. 3(11): p.917-22.

Tech ID/UC Case

19789/2005-186-0

Related Cases

2005-186-0

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