CRISPR Arrives at ICBR

CRISPR at UF ICBR

In collaboration with the UFHCC, ICBR is pleased to announce several new CRISPR genome editing services now available to the UF research community.

The CRISPR/Cas9 system allows researchers to easily introduce deletions, insertions and edited DNA sequences into mammalian cell genomes. It can be used to introduce very specific surgical edits to any gene and can in fact introduce foreign genes into specific regions of the animal cell genome.

At the University of Florida, CRISPR is used within research laboratories by principal investigators with the required technical background in molecular biology. However, researchers without the necessary experience to perform CRISPR are reliant on either collaborations with other researchers, or commercial entities that provide CRISPR services. For many, the use of vendors is cost-prohibitive, while the busy schedule of UF PIs can often hinder the formation of collaborations with fellow faculty members who need assistance with CRISPR-based protocols.  To ensure all UF researchers have access to this valuable gene editing technology, the ICBR and UFHCC have collaborated to from a CRISPR Technology Core as part of the ICBR Gene Expression and Genotyping Core. This new resource provides services for generated mammalian cell lines in which one or both copies of a particular gene have been altered by the introduction of random mutations. The CTC is under the technical direction of UFHCC member Dr. Chris Vulpe, Professor in the Department of Physiology and resident CRISPR technology expert.

According to ICBR Director Dr. Rob Ferl the CTC “meets the need for a cost efficient gene editing core here on campus and exemplifies the value of collaborative approaches to moving science forward on the UF campus”.

UF researchers interested in gene editing can arrange for a free consultation with Dr. Vulpe who will then design the best approach for attaining the desired animal or plant cell gene mutation. Dr. Vulpe will also provide consultation on how to use CRISPR technology to conduct for phenotypic screens in mammalian cells. If you are interested in learning more about the CTC, contact Dr. Vulpe (cvulpe@ufl.edu) or Dr. Steven Madore (smadore@ufl.edu), ICBR Associate Director for Science. Additional information on CRISPR can also be found on the ICBR web site under the Gene Expression and Genotyping Core.

What is CRISPR?

CRISPR, an abbreviation for Clustered Regularly Interspaced Short Palindromic Repeats, is actually a family of short, foreign DNA sequences that were discovered in bacteria over 30 years ago. The sequences are copied from invading viruses and then inserted into the bacterial genome, and are used by the bacteria as part of an immune defense system to destroy DNA detected from the similar viruses during subsequent infection. The short CRISPR DNA sequences are converted into a small RNA molecule that when combined with a bacterial enzyme that cuts the DNA, recognizes invading viral DNA and targets it for enzymatic destruction. Recently, molecular biologists have shown that by combining small CRISPR-like RNA molecules designed to recognize specific gene regions with the bacterial enzyme Cas9, animal and plant gene can be targeted for cutting. This gene editing machine is referred to as the CRISPR/Cas9 system. Lesions in the CAs9-cut DNA are rapidly repaired by existing DNA repair mechanisms present and on alert in all cells. However, the repair process is not always perfect, such that a significant portion of the cut gene regions will be repaired incorrectly, resulting in genes that now have deletions or insertions in the DNA coding region. These mutations can have deleterious effects on the “edited” gene such that the protein it encodes in dysfunctional in some way. In addition to introducing random insertions or deletions into animal cell genes, the CRISPR/Cas9 system can be used to introduce very specific surgical edits to any gene, and can in fact introduce foreign genes into specific regions of the animal cell genome. Thus, CRISPR gene editing now allows researchers to easily introduce deletions, insertions, and edited DNA sequences into mammalian cell genomes.