CRISPR in a Box™ Educational Kits

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CRISPR in a Box™ Educational Kits

In partnership with the world-renowned gene editing team at ChristianaCare and the experienced instructors from Delaware Technical Community College, Rockland is proud to provide these revolutionary educational kits to classrooms and life science companies around the world.

Because of the efforts of these partners and the support from the National Science Foundation, these kits have already been used to train more than 100 students and over 50 community college faculty across the US—while being modified and improved to keep pace with the ever-expanding technology. Now, we aim to make them globally accessible, training the next generation of scientists and ultimately advancing life science to foster a better world.

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Advantages of CRISPR in a Box™ 

The CRISPR in a Box™ Educational Kits utilize 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 kits were 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.

no sophisticated equipment

No sophisticated equipment required

cell free reactions

Safe, cell-free reactions

adaptable to any curriculum

Adaptable to any curriculum

With CRISPR in a Box, you'll be able to:

  • Understand how CRISPR interacts with DNA
  • Carry out gene editing reactions in a cell-free environment
  • Visualize all gene editing products from a single reaction
  • Develop a fundamental understanding of the degree of mutagenesis and precise or error-prone gene editing outcomes
  • Explore the effects of different repair pathways acting simultaneously (HDR, NHEJ, MMEJ, etc.)
  • Study gene knockout and knock-in strategies in one reaction

 

How CRISPR in a Box™ works

The model system uses CRISPR-Cas12a ribonucleoprotein (RNP) in vitro to modify a segment of the lacZ gene with 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 then transformed into Escherichia coli (E. coli) bacteria, producing a visual, phenotypic readout of a blue-to-white color change.

 

An Overview of the CRISPR/Cas System

CRISPR or Clustered Regularly Interspaced Short Palindromic Repeats is a groundbreaking technology that can harness the activity of DNA repair pathways to generate desired genomic alterations. The development of site-specific CRISPR-directed gene editing technologies has provided invaluable tools to examine structure-function relationships of eukaryotic genes. This revolutionary tool is currently providing novel therapeutic modalities for the treatment of inherited diseases and cancer, accelerating drug discovery, and creating a brand-new class of diagnostics. The extraordinary versatility of CRISPR for gene modification and genetic detection will undoubtably lead to the development of applications that have yet to be conceived. It will be the next generation of scientists who will continue to uncover these novel applications and take advantage of the exceptional potential of this molecular machine.

CRISPR initiates double-stranded cleavage of DNA at specific target sites, triggering DNA double-stranded break (DSB) repair. Generally, DNA repair occurs through two distinct pathways: error-prone non-homologous end joining (NHEJ) or precise homology directed repair (HDR). As an initial response, and if no suitable repair template is present, the cell attempts to repair the damage through NHEJ. During the NHEJ response, DNA ends are processed via ends being joined back together. This often results in disruptive DNA insertions and/or deletions (indels) occurring at the break site. HDR is a broad classification of repair pathways requiring a homologous donor template to direct a slower, more precise process of repair which relies on a series of reparative steps capable of recapitulating either the endogenous sequence or resulting in mutagenesis. The alignment of an appropriate donor repair template with homologous regions around the resected ends of the DSB site provides a guide to drive the cellular machinery to facilitate a specific repair outcome.

 

CRISPR in a Box™ Kit Details

Starter Kit
CRISPR in a Box Starter Kit

✓ Cas12a protein

✓ crRNA1364

✓ Cell-free extract

✓ HDR-NS Oligonucleotide (ODN)

 

 

 


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Complete Gene Editing Laboratory Kit
CRISPR in a Box Complete Kit

✓ Cas12a protein

✓ crRNA1364

✓ Cell-free extract

✓ HDR-NS Oligonucleotide (ODN)

✓ All required buffers

✓ All required enzymes

✓ Petri dishes, tubes, and more!


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Other materials needed but not provided:

  • 2–20 µL Adjustable Micropipet and Tips
  • 20–200 µL Adjustable Micropipet and Tips
  • 200–1000 µL Adjustable Micropipet and Tips
  • Autoclave or Microwave Oven
  • Temperature-controlled Dry Bath/Water Bath
  • Benchtop Centrifuge
  • Soaking Incubator
  • Horizontal Gel Electrophoresis Chamber with Gel Casting Tray and Comb
  • Power Supply
  • Blue Light or UV Transilluminator
  • Autoclavable Bottle with Cap
  • Erlenmeyer Flask
  • Graduated Cylinder
  • Deionized Water
  • Wet Ice and Container (i.e. ice bucket)
  • Permanent Marking Pens
  • Tube Racks
 

Expected Results & Readouts

Figure 2. Restriction enzyme digestion on an agarose gel
Figure 3. Gene editing reaction bacterial plates

FAQs


  1. How many kits/reactions will I need for my class?
    The kit has 10 reactions. We recommend that students work in pairs or individually; however, three students/group is also possible.

  2. What skill level is required to use this kit?
    Basic laboratory skills in sterile technique and micropipetting is required.

  3. My students are not very precise yet, will they achieve positive results?
    Yes. Because of the robust nature of this kit, positive readings are expected on the majority of final plates even if subtle pipetting errors occur.

  4. My lab period is only 1 hour, will this kit be possible?
    Yes, the gene editing reaction can be further broken down into ~1 hour segments, storing the products at –20ºC until the next class.

  5. Do you need to sequence the DNA to see the outcomes?
    No, there is a phenotypic bacterial colony color change from blue to white. To show students a genotypic result, a restriction enzyme digestion may be done on colonies to detect which colonies were edited to contain the NotI insertion.

  6. I have my own competent cells, will these work instead of the recommended DH5α cells?
    Likely yes, as long as they exhibit a comparable genotype. However, a transformation reaction will need to be tested before using with the class. Many competent cells require an optimal dilution when plating, but we have not tested every competent cell on the market.

  7. I don’t have an incubator; can the cells grow at room temperature?
    While the timing will be much slower than in an incubator, the cells should be able to grow at room temperature although it is not recommended.

  8. What class can this kit be incorporated into?
    We believe that this can be done in microbiology and general biology classes where work with bacteria is a common practice. This kit may also be well suited for higher-level core class like genetics, molecular and cell biology classes because of the practicality and usage in field science and healthcare.