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Running Head: VirologyVIROLOGY
Virology2Aims and objectivesSeveral stringent cellular and molecular techniques have been used to identify and portray the prohibitin-2as a receptor of infection of Dengue in a bug. In order to study the virus namely the Host interaction andthereby proving that PHB-2 is a receptor of Dengue virus in the cells of the insects CRISPR/Cas9 is beingused to knockout the prohibitin-2 gene. Likewise in order to manipulate the PHB-2 gene, which wouldresult in the mutation of genomic sequence, there has been an aim to prove that PHB-2 gene could also besilenced just to stop the functional roles with the help of CRISPR/Cas9 tools. Bi-allelic mutation musthappen to accomplish the effective gene function silencing. For stopping the virus namely Dengue to enterthe insect cells will definitely silence the PHB-2. Thus, the infection of dengue would be reasonablebecause of controlling of the vector being spread.Research Design Overview:Dr. Shin-Wan Chan was the person to start this project, and this project is for the characterization as wellas identification with the help of few techniques. For providing an overview of the project workflow, thestudy has been designed in three parts namely- parts done by Dr. Shiu-Wan and few of the stuents pursuingMasters. The part one of the study includes- Firstly, plasmid vector namely (E-coli) cloned into SgRNA.Puromycin resistance gene is the vector that is encoded Cas9 which act as a selectable marker for theindication of transfection. On the second hand, Using lipid by the transfection of plasmid vector. Thesehelp in the continuous growth and helps the cells maintain the selection pressure thereby making thepuromycin in the growth medium. Puromycin helps in the killing of large quantity of cells therebyindicating that the plasmid is lost with puromycin resistance gene, this takes places after one to two weeksof the first step. Retention of the plasmid is through the cells that remain growing in the puromycincontaining medium and this may have stably integrated into the genome of the targeted cells.