CRISPR/Cas9 design to knockout and knockin the breast cancer gene-BRCA1 in Arabidopsis thaliana

Abstract

Arabisopsis thaliana is a model for many molecular and genetics research especially in human research, there are many genes in A. thaliana sharing them with human. BRCA1 is a gene related with breast cancer, any damage in it sequences may leads to raise the percentage of cancer tumor. AtBCRA1 a breast cancer gene in A. thaliana plant were obtain and blast from NCBI web tool and used for gene editing, a set of PCR primers were designed to detect this gene by using in- silico tools and simulate the PCR reaction and gel electrophoresis. As well as a gene editing tool, CRISPR/Cas9 were designed to knockout and knockin any mutation in the BRCA1 gene by designing sgRNA as a target site in DNA for CRISPR action with a set of primer to amplifying the target by using CHOPCHOP web tool. A good target sequences design is a very critical step in CRISPR/Cas9 technology success.