1: You have been tasked to perform a CRISPR based knockout of your gene. Identify all candidate sgRNAs which can knockout all isoforms of your gene. Paste the excel spreadsheet with a list below.

 

 

2: Annotate in benchling where these candidate sgRNAs are

 

3: For two of the sgRNAs you have found, design PCR primers which will amplify the target site and produces a product less < 1000 bp

 

4: Indicate where in the target site the double strand break will happen.`

 

5: What is the impact on the protein coding sequence if the following NHEJ mutations occur:

(A) Single base deletion

(B) Two base deletion

(C) Three base deletion


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