At the height of the COVID-19 pandemic in 2020, Marvin Collins ’22, a bioengineering student, was balancing their Stanford classes from home in Alabama while also helping bioengineering professor Stanly Qi create a CRISPR/Cas9 gene-editing kit from a makeshift lab in their parents’ garage. They lacked typical lab equipment or reagent storage, evening having to buy a $30 chicken egg incubator from Amazon to control temperature over the course of the experiments.
Fast forward five years to January, and the kit, named “CRISPRKit,” is now being disseminated through a pilot program to 500 high schoolers across 25 high schools across the Bay Area.
Collins, along with Matthew Lau ’25, developed the affordable CRISPRKit to make CRISPR gene-editing technology accessible to high school students underrepresented in the sciences. The kit was created during their time as undergraduates under the mentorship of Qi, a CRISPR/Cas9 researcher. Qi believes that college students are critical in making biotechnologies within the life sciences more understandable and accessible for the general public.
“Universities are so good at making cutting-edge technologies, but sometimes they can be out of people’s scope,” Qi said.
For Lau, the broader purpose of the kit is to “democratize biology,” especially with CRISPR/Cas9 being a significant “black box of biology” for many.
James Stiltner, a biotechnology teacher at Los Altos High School, tested CRISPRKit with his students. He said CRISPR technology applications hold great promise to be a focal point in society for coming generations, underscoring CRISPRKit’s relevance to today’s youth.
CRISPR/Cas9 kits provide miniature experimental set-ups that allow individual students to visualize gene-editing, typically through fluorescent color changes. Existing CRISPR/Cas9 gene editing kits currently cost at least $100 per student, on top of expensive lab equipment, according to Qi. The newly developed CRISPRKit costs about $2 per student and can be used in a typical high school classroom with only a smartphone.
Tanya Buxton, an Advanced Placement biology teacher at Menlo School, also taught students who tested CRISPRKit, which she said has an “elegant and simple” design. According to Buxton, CRISPRKit provided her students an opportunity for “high level learning” by allowing them to visualize gene-editing “with the naked eye.”
In a Nature Communications study published in August, CRISPRKit was tested on seventy five high school students from three different Bay Area high schools. Collins and Lau were co-first authors of the paper.
The team has received almost 8,000 kit requests internationally, including from the United Kingdom, Germany, the Philippines and Brazil. The team also hopes that their pilot program will prove to manufacturers that their kit is “viable on a broad scale” and ready for distribution statewide, especially to Title I high schools in California.
The CRISPR/Cas9 gene-editing system, based on a bacterial immune response against viruses, has been applied to human genome editing and was recently implemented in the first Food and Drug Administration-approved gene-editing drug Casgevy, which treats sickle cell anemia, a blood disorder.
CRISPR/Cas9 works using a Cas9 enzyme and a guide RNA. Lau compared the guide RNA to a global positioning system (GPS) that scans the genome and leads the Cas9 to cut DNA at a specific sequence, which can activate or deactivate specific genes.
CRISPRKit works in a similar way, though it employs dCas9, or “dead” Cas9, a deactivated version of the Cas9 enzyme unable to cut DNA.
“Whereas CRISPR-Cas9 is normally a pair of molecular scissors, dCas9 acts as an on-off switch,” Qi said.
The dCas9 enzyme sticks to specific DNA sequences and blocks protein production and gene expression. According to Qi, the fact that this method lacks any direct DNA editing makes it well-suited for the classroom. The Qi Lab is the first lab to develop a working dCas9 protein.
CRISPRKit experiments are carried out within a cell-free system, which contains the contents of cells that were previously broken open. These mixtures, which create artificial cell-like environments for protein production and gene-editing, are now being supplied by the Jewett Lab, a bioengineering lab at Stanford that specializes in cell-free synthetic biology, Lau said.
According to Collins, these cells can’t reproduce in the same way as normal cells living in bacteria, but they can still perform the same essential processes involved in making proteins.
“We wanted to make the readout interesting, straightforward and fun for students using colors,” Qi said. To align with current high school curricula, especially AP biology, the team plans to move forward with a version of their kit that leverages dCas9 to repress the production of melanin, which is responsible for hair and skin color.
According to Lau, CRISPRKit demonstrates how various guide RNAs can repress genes with differing levels of effectiveness. The team designed the melanin CRISPRKit to include guide RNAs leading dCas9 to different regions of the gene for the tyrosinase enzyme, which is normally involved in the conversion of an amino acid into melanin.
The experiments themselves take thirty minutes and after a day of incubation, students can analyze color readouts to predict which guide RNAs were the most effective repressors by taking a picture with their smartphone and uploading it to the CRISPectra website.
“The greater the success of your gene switch, the more decrease in color you see,” Qi said.
According to Collins, the COVID-19 pandemic informed the team’s approach to CRISPRKit by “highlighting an existing need” for accessible STEM education. Despite CRISPR becoming a scientific breakthrough and a significant topic of interest, gene editing technologies can be outside of peoples’ scope.
“It requires a lot of innovation, creativity, and effort but college students could have a profound impact democratizing broader biotechnology, biology or life sciences so that their parents, grandparents and the broader general public can better understand them,” Qi said.