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Related Concept Videos

The Ras Gene02:38

The Ras Gene

The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a superfamily...
The Ras Gene02:38

The Ras Gene

The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a superfamily...
Rous Sarcoma Virus (RSV) and Cancer01:03

Rous Sarcoma Virus (RSV) and Cancer

Rous Sarcoma virus or RSV was discovered by F. Peyton Rous in the year 1911 as a filterable transmissible agent that could cause tumors in chickens. He won a Nobel Prize for this discovery in 1966. His experiments clearly demonstrated that some cancers could be caused by infectious agents and led to the discovery of many more cancer-causing viruses in animals as well as humans.
RSV is a retrovirus that contains two copies of a plus-strand  RNA genome. Its genome consists of four main open...
Abnormal Proliferation02:23

Abnormal Proliferation

Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the daughter...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

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CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion
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Construction of chimeric E3s expression plasmids targeting oncoprotein ras.

Yi-hui Ma1, Qiang Zhang, Yu-mei Gu

  • 1Department of Pathology, PUMC Hospital, CAMS and PUMC, Beijing, China.

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. Acta Academiae Medicinae Sinicae
|September 8, 2012
PubMed
Summary
This summary is machine-generated.

Researchers developed novel chimeric E3 expression plasmids to target and reduce oncoprotein Ras levels. This protein knockdown strategy offers a new avenue for cancer research and therapeutic development.

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Area of Science:

  • Molecular Biology
  • Oncology
  • Biotechnology

Background:

  • Oncoprotein Ras plays a critical role in various cancers.
  • Targeting Ras for protein knockdown is a promising therapeutic strategy.
  • Developing efficient expression systems for protein knockdown is essential.

Purpose of the Study:

  • To construct chimeric E3 ubiquitin ligase (E3) expression plasmids targeting oncoprotein Ras.
  • To utilize protein knockdown theory for Ras inhibition.
  • To create a foundation for further research in Ras-targeted therapies.

Main Methods:

  • Chimeric E3 plasmids were engineered using specific binding and functional domains (Raf-1, PI3K, RalGDS, F-Box, U-Box).
  • Plasmid integrity and nucleotide fragment cloning were validated using double enzyme digestion, PCR, and sequencing.
  • Expression efficiency in eukaryotic cells (HEK293T) was assessed via Western blot analysis.

Main Results:

  • Five out of six constructed plasmids successfully expressed fusion proteins in HEK293T cells.
  • The plasmid (RBD+CRD)(Raf-1)-U-Box-pcDNA3.1 demonstrated the ability to effectively knock down Ras protein levels in PANC-1 cells.

Conclusions:

  • Novel chimeric E3 expression plasmids targeting Ras were successfully constructed.
  • These plasmids provide a robust platform for investigating protein knockdown strategies against Ras.
  • This research lays the groundwork for developing new cancer treatments focused on Ras inhibition.