I want to knock in GFP to my interested geen, i want to use cas9 to acheive it.i konw the pricipal,but i did not konw the experiment detailed of it??

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Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

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Sinan Kilic

Hi Pei Cai

There is a quite detailed protocol here using dual nickases and explaining how to design and test your Cas9 and your cells after editing:

Article Generation and validation of homozygous fluorescent knock-in...

Also keep in mind some of the things discussed in this thread about endogenous protein levels and choice of fluorescent protein:

https://www.researchgate.net/post/How_to_improve_the_expression_of_fluorescent_protein

Pei Cai

Sinan Kilic thank you so much! I will read your recommended paper and link carefully. Your answer make me so excited for my first time ask for help on "researchgate "and you replied me. Thank you again