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