Publication: Construction Of A CRISPR Based Cloning Vector Targeting Ace2
Loading...
Date
2023-09
Authors
Gunasekaran, Thanes
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
COVID-19, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in late 2019, poses a significant threat to public health and safety. Efforts to manage the pandemic, including immunizations, have been limited by the emergence of novel SAR CoV-2 variants andthe erosion of immunity. The Protein-Protein interaction network for ACE2 was constructed using Cytoscape 3.10.0 to study the interaction of ACE2 with other genes or proteins involved in the pathogenesis pathway of SARS-Co V -2. Analysis of the ACE2 network showed interaction with nine genes (ACE2, TMPRSS2, CLEC4M, DPP4, ADAM 17, ANEP, CCL2, NPCl, & MEPLA) involved in virus infection process. Besides that, the gene expression analysis of ACE2, TMPRSS2, Arg 1 & IL6 was determined using RT-PCR. The analysis indicated that ACE2 had the highest expression level compared to other three genes in HEK 293 cell line and provided a better understanding of expression of ACE2 in kidney cell line and its related signaling pathway. Apart from that, the construction of an ACE2-knockout CRISPR ·plasmid was successfully executed by designing gRNA and cloning it into a CRISPR plasmid, pX458.
Description
Keywords
Thanes , Gunasekaran , Construction Of A CRISPR , Based Cloning Vector Targeting Ace2