Silencing the AGOS7 gene ın tobacco (Nicotiana tabacum) plant using the CRISPR/CAS9 system

Tobacco (Nicootiana tabacum L.) is an annual plant of Solanaceae family and categorized in delightful plants. Economically important tobacco is also classified as an industrial and aromatic plant. Tobacco which can adapt to diffirences of geographic conditions quickly and efficiently is known as one of the agricultural plants which have best ability to adapt. DNA-free genome editing can reduce some limitations. These methods allow us to create gene specific mutations to increase resistance against biotic stress factors. Additionally, they can be used to demonstrate the functions the genes. Consequently, modern genome editing tecnics such as ZFN, TALEN CRISPR/Cas9 apeared as powerful genome editing tools. Because of its low cost and applicability, CRISPR/Cas9 has become most popular genome editing tool in the last 8 years. Proteins of Argonaute gene family is one of the most important protein families of plants. Argonaute gene family can be found in many species and has very important role in RNA-medieted gene silencing. Proteins of this gene family has two domains (PAZ and PIWI) which have critic roles in stress respoonse and development. Previous studies demonstrated that there are diffirence in gene expression of AGO genes under abiotic stress such as drought, salinity, high temperature and, low temperature. Moreover, there are some studies showed funtions of AGO genes in cell expansion, cytoskeleton and, programmed cell death. AGO7 (LOC107784577) is chosen based on homology of other model organisms, and knocked-out using CRISPR/Cas9. The results showed that there are diffirences in morphology and physiology between AGO7 and wild-type plants.