Understand how CRISPR interacts with DNA using this innovative, educational gene-editing activity.
- Safe, cell-free reactions
- Adaptable to full curriculum
The CRISPR-In-A-Box™ Kit utilizes revolutionary CRISPR-Cas technology through an innovative educational activity that expands the understanding of this powerful genetic tool. The hands-on process introduces students to invaluable genomic engineering techniques that can impact future therapy in drug discovery and diagnostics. The kit was specifically designed to show the entire spectrum of gene editing products that occur in a CRISPR/Cas reaction. And unlike its competitors, the basis of the laboratory exercise is done on a plasmid instead of a functional gene in bacteria, generating a much safer reaction. Including prep, the complete lab will take approximately 3 days to 1 week to complete.
The model system provided uses CRISPR-Cas12a ribonucleoprotein (RNP) in vitro to modify a segment within the lacZ gene using a single-stranded donor DNA template and a mammalian cell-free extract. The single-stranded donor DNA template contains flanking arms of homology to the target site and directs the integration of a NotI restriction enzyme site. The products of this reaction are transformed into Escherichia coli (E. coli) bacteria and a visual, phenotypic readout of a blue to white color change is seen. Additional downstream sequencing or restriction enzyme digestion can be used to confirm presence of the integrated NotI site. One of the many advantages of the in vitro system is the ability to analyze all gene editing products from a single reaction. Other major advantages of utilizing this system include, but are not limited to, exploring different repair pathways (HDR, NHEJ, etc.), targeting a variety of unique genes, and evaluating the efficiency of diverse Cas proteins.
Ordering information: Materials and equipment required but not included: adjustable volume micropipettes (2-20, 20-200, 200-1000uL) and corresponding tips, adjustable bottles with caps (250ml and 1L), autoclave, microwave oven, benchtop centrifuge, microcentrifuge, blue light or UV transilluminator, refrigerator, -20 freezer, -80 freezer, DI water, Erlenmeyer flasks (125ml, 250ml), gloves, graduated cylinders (100ml, 500ml, 1L), horizontal electrophoresis chamber with gel casting tray and comb, power supply, shaking incubator, spectrophotometer and associated materials, temperature-controlled dry bath or water bath, tube racks, ice.
Delivery information: The Starter Kit contains the Cas12a protein, crRNA1364, cell-free extract, and HDR oligonucleotide. The Complete Gene Editing Laboratory Kit contains all of the items from the Starter Kit as well as all required buffers and enzymes, petri dishes, tubes, and other reagents.
