Step 1: Prepare input file

CRISPR-BETS allows Genbank, Snapgene, Fasta(sequence) file formats as input.

 

Step 2: Select and upload file

  • ① upload GenBank or Snapgene file

    1. Navigate to the Select load file type panel. click GenBank button (GenBank is the default input file).

    2. Upload the GenBank file.

    3. Select gene isoform (support for Genbank and Snapgene file).

  • 1. Navigate to the Select load file type panel. click Fasta(sequence) button

    2. Fill the text box with both the DNA and CDS sequences of a gene.

    NOTE: In order to design cross-CDS gRNA, enter both DNA and CDS FASTA files or sequences. Currently, only one gene is supported by an analysis.

Step 3: Select edit system option

1. Navigate to the Edit system option panel.

2. (optional) Select Cas9 variants, PAM and gRNA length will be automatically filled.

3. (optional) Select Deaminase, Edit windowwill be automatically filled.

4. The user fill PAM, gRNA length and Edit window.

5. (optional) The user select genome to estimate the specificity of each gRNA.

6. Click the Analysis button and wait for the analysis to complete.


Parameter description

  • 01 Cas9 variants

    Select the appropriate Cas9 variant. Once selected, the parameters PAM and gRNA length will be filled in automatically, not required.

  • 02 Deaminase

    Select the Deaminase. Once selected, the parameters Edit window will be filled in automatically, not required.

  • The default value is NGG, which can be customized, required.

  • The default values are 1 and 20 (The farthest first base of PAM is represented as a 1), which can be customized, required.

  • The default value is 20, which can be customized, required.

  • Select species genome for off-target analysis, this step will increase the running time of CRISPR-BETS, not required.

 

Step 4: Scanning result panel and download result information

1. Click the button and wait for the analysis to complete.

2. Navigate to the result panel, the user can drag and zoom to check the detailed information of each gRNA.

3. Click result as txt button to download result.


The following two functional buttons exist in the upper right corner of the results panel



Output result description