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CRISPR Cas 9 Gene Editing

CRISPR Gene Editing Plasmid Design Bioinformatics Genetics

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#1 Salma Sweeti

Salma Sweeti

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Posted 30 July 2022 - 03:49 AM

I am trying to design a plasmid for a knock in experiment and wanted to clarify some technical issues. 
 
I wanted to know exactly which components must be included in the plasmid. 
 
I know that I have to include the 5' homology arm , insertion gene sequence, marker sequence and 3' homology arm. 
But, I would like to know for certain the following details:
 
1) Should I take the whole sequence of the gene or do I need to remove the introns/UTR regions or any other unnecessary part of the sequence?
2) Which sequence should I choose for gene insertion , gene sequence or mRNA transcript sequence?
3) When designing plasmids, should I insert extra promoters and/or terminators with the insertion genes?
 
Additionally, can you recommend an online tool to design HDR Plasmid? 






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