1. GRNA DESIGN
Faculty of Advanced Technologies of Medical Sciences
Reza Rakhshi
Master student of medical biotechnology
2. CHOPCHOP SITE:
REZA RAKHSHI, MASTER STUDENT OF MEDICAL BIOTECHNOLOGY
RefSeq/ENSEMBL/gene name
or genomic coordinates.
Choose an Organism
3. CHOPCHOP SITE:
REZA RAKHSHI, MASTER STUDENT OF MEDICAL BIOTECHNOLOGY
Change default PAM and
guide length in Options.
Choose your goal
4. CHOPCHOP SITE- OPTIONS:
REZA RAKHSHI, MASTER STUDENT OF MEDICAL BIOTECHNOLOGY
Better than we target CDS or First and
Second exon
Optimum range for GC content is
40-60%
Self complementary is ZERO
5. CHOPCHOP SITE- OPTIONS:
REZA RAKHSHI, MASTER STUDENT OF MEDICAL BIOTECHNOLOGY
Optimum range is between 19-20
Depends on Cas choose best sequence
Optimum range for first parameter is 3
And for second parameter is ZERO
G20 so important to us
Depends on our cell line
GG or NG/GN important for my RNA
Polymerase (T7 and U6)
Finally PRESS
“ find the target site”
6. RESULT:
REZA RAKHSHI, MASTER STUDENT OF MEDICAL BIOTECHNOLOGY
Analyze results and
choose the best.
1. PAM seq.
2. G20
3. The 5′
base
4. Repeated base
7. DATA ANALYZE:
REZA RAKHSHI, MASTER STUDENT OF MEDICAL BIOTECHNOLOGY
Low mismatch in 3′
is
better than
Possibility of
framshift
Possibility of
deletion
8. CAS-OFFINDER- PRECISE ANALYSIS OF MISMATCH:
REZA RAKHSHI, MASTER STUDENT OF MEDICAL BIOTECHNOLOGY
Choose your PAM seq
Write your gRNA seq.
without PAM seq.
Respectively
Write:
3
1
1
finally
