Dear All,
I want to design sgRNA target sequence and insert into scaffold portion already present in pRGEB31 binary vector. Any experimental protocol for that?
I want to knock out a disease susceptible gene in citrus. Is there any specific database to get the complete genome sequence of sweet orange (Citrus sinensis)? So that i can choose that target gene sequence to design sgRNA and clone it into the above-mentioned vector. Thanks in advance.
Please find the below information
Useful sites for sgRNA design
• http://www.genome-engineering.org/crispr/?page_id=41
• http://www.addgene.org/CRISPR/guide/
• http://crispr.mit.edu/
To find predesigned sets of sgRNAs (see example for mouse RAD51):
• http://genome.ucsc.edu/cgi-bin/hgTracks?db=mm9&position=chr2%3A118933851-118949804&hgsid=369022555_n8aBpwayTCGjo561eWeJ8Z9QnyqL
please look over here: https://www.neb.com/-/media/nebus/files/application-notes/construction-of-an-sgrna-cas9-expression-vector-via-single-stranded-dna-oligo-bridging-of-double-stranded-dna-fragments.pdf
Article Insert, remove or replace: A highly advanced genome editing ...
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