Generation of Genomic Deletions (of Rig-I GENE) in Goat Primary Cell Culture Using CRISPR/CAS9 Method.
Shivani MalpotraAshutosh VatsSushil KumarDevika GautamSachinandan DePublished in: Animal biotechnology (2017)
CRISPR/Cas9 system is a natural immune system in prokaryotes protecting them from infectious viral or plasmid DNA invading the cells. This RNA-guided system can act as powerful tool for introducing genomic alterations in eukaryotic cells with high efficiency. In the present study, Rig-Igene is taken as model gene to study the efficiency of CRISPR/Cas9 system induced gene deletion in primary fibroblast cell culture. Rig-I(retinoic acid-inducible gene-1) is involved in regulating immune response in mammals. In this study, we optimized the CRISPR/Cas9 method for knocking out Rig-Igene in Goat primary fibroblasts by using a NHEJ pathway. Cells were screened for inactivation of the Rig-Igene and two positive clones were found out of thirty colonies screened. Thus, cells containing Rig-Igene inactivation could be achieved by CRISPR/Cas9 in goat fibroblast cells.
Keyphrases
- crispr cas
- induced apoptosis
- genome editing
- cell cycle arrest
- immune response
- copy number
- genome wide
- endoplasmic reticulum stress
- high efficiency
- escherichia coli
- oxidative stress
- gene expression
- toll like receptor
- dendritic cells
- inflammatory response
- extracellular matrix
- drug induced
- stress induced
- circulating tumor cells