2024 NSTA New Orleans • EDVOTEK® Workshops

06 - Introducing Your Students to CRISPR with Sickle Cell Gene Editing

Experimental Results and Analysis, continued

MODULE II A representative gel can be seen below. DNA samples in lanes 3 and 5 show two bands, indicat- ing that the DNA has been digested. This indicates that gRNAs #2 and #4 were successfully able to target the DNA for cleavage by Cas9.

----- Did not target Targeted DNA Did not target Targeted DNA Did not target

----- 4300 bp 3000 bp/1300 bp 4300 bp 3000 bp/1300 bp 4300 bp

Standard DNA Marker gRNA #1 gRNA #2 gRNA #3 gRNA #4 gRNA #5

1 2 3 4 5 6

Based on these results, guide RNAs #2 and #4 seem to be the best candidates to target the BCL11A gene. At this point, testing could continue to determine whether the CRISPR:Cas9 com- plex and this gRNA could be used to digest the target DNA in cells.