You design a guide RNA to target your gene of interest, which you will express attached to a scaffold RNA. You transform cells with Cas9, the guide+scaffold RNA, and an HDR template. The HDR template codes for the insertion of a premature stop codon located 10 bp upstream of the Cas9 cut site, a base pair substitution 15 bp upstream of the Cas9 cut site, and a base pair substitution in the PAM sequence. Question: Is this scenario “possible” or “not possible”? Cas9 cuts the target gene sequence at a site 3 bp downstream of a PAM site Explain your reasoning as to why
Gene Interactions
When the expression of a single trait is influenced by two or more different non-allelic genes, it is termed as genetic interaction. According to Mendel's law of inheritance, each gene functions in its own way and does not depend on the function of another gene, i.e., a single gene controls each of seven characteristics considered, but the complex contribution of many different genes determine many traits of an organism.
Gene Expression
Gene expression is a process by which the instructions present in deoxyribonucleic acid (DNA) are converted into useful molecules such as proteins, and functional messenger ribonucleic (mRNA) molecules in the case of non-protein-coding genes.
You design a guide RNA to target your gene of interest, which you will express attached to a scaffold RNA. You transform cells with Cas9, the guide+scaffold RNA, and an HDR template. The HDR template codes for the insertion of a premature stop codon located 10 bp upstream of the Cas9 cut site, a base pair substitution 15 bp upstream of the Cas9 cut site, and a base pair substitution in the PAM sequence.
Question: Is this scenario “possible” or “not possible”?
Cas9 cuts the target gene sequence at a site 3 bp downstream of a PAM site
Explain your reasoning as to why.
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