Clustered regularly interspaced short palindromic repeats or CRISPRs are locations on DNA containing short base repetitions. These locations arise in bacteria as a result of exposure to a virus or foreign pathogen. These regions are able to attract a variety of cas genes that code for proteins related to the CRISPR region. The end result is the generation of a targeted nuclease that is able to silence the foreign virus/DNA at the DNA or RNA level.
The CRISPR system demonstrated an efficient method harnessed by bacteria to target and cut specific regions of DNA. These qualities made the CRISPR/CAS system a logical choice for genome editing. Additionally the enzyme responsible for cutting DNA, Cas9, had successfully been altered to cut a single strand of DNA without affecting its targeting mechanism. In 2012 CRISPR was successfully used as a genome editing tool in a variety of species.
Through the use of a combination of crRNA and tracrRNA (the cas generated guiding components in native CRISPR) called sgRNA, Cas9 can be targeted to a specific genomic region. Once attached the enzyme can make a single stranded ‘nick’ or double stranded ‘break’ at a specified nucleotide. This region can be altered through the addition, subtraction or alteration of the sequence. Generally these elements are added using plasmids that must be integrated into the host cell’s DNA. Allele’s RNA based CRISPR platform harnesses modified RNA to introduce these elements exogenously (footprint-free). Additionally RNA allows for highly efficient expression that can be tightly controlled through dosage.
Our RNA based CRISPR service offers a number of advantages over traditional genome editing approaches. The RNA platform allows for highly efficient 'footprint-free' genome editing that is precisely controlled through RNA dosing. This precise controll couple with highly precise sgRNA design allows for unmatched expression and the elimination of off target effects. Additionally, our extensive experience in the field of cellular reprogramming gives you the option to start with iPSCs or any iPSC derived cell types, potentially providing you with a limitless source of material. Lastly, our RNA-CRISPR services harness the latest deep sequencing technologies to ensure only the desired alterations are made.
Phase 1- Choose your cell type (iPSC's, Fibroblasts, etc.) and desired modifications. Your experiment will be analyzed and designed by our scientific team.
Phase 2 - Your experiment will be carried out.
Phase 3 - The resulting cells will be analyzed using next generation deep sequencing technologies to ensure only the desired modification were made.
Phase 4 - Successfully edited cells can be modified or integrated into various cellular assays.
Every genome modification project is custom in nature, to find out more about our serivces or request a quote please fill out our custom service contact form.
mRNA based CRISPR/Cas9 genome editing offers a number of advantages over traditional plasmid based CRISPR. mRNA is non-integrating, allows for a high degree of expression controll, and efficient gene expression. Below is our current list of CRISPR/Cas9 mRNA.
We will update this list as we release more products for this platform.