Most biologists are familiar with the precision gene editing system CRISPR. But what about SNIPER†? SNIPER is a digital PCR (dPCR) and culture-based screening technology that has the power to enhance CRISPR-Cas9 gene editing, enabling multiplex gene modification, biallelic gene insertions and increased screening efficiency. Read on to find out more about how SNIPER can enhance your precision gene editing protocols and save you time, money and effort.
1. Insert large genes
Using the dPCR-based SNIPER screen, it is easier to detect large gene fragments edited by CRISPR technology, such as selection or reporter genes. Using conventional screening methods, the detection rate declines when insert sequence size exceeds 2 kbp. However, SNIPER can detect genes up to 7 kbp in size.
The improved sensitivity and specificity of this advanced screening technique can achieve up to 30 times* the detection rate compared with conventional screening methods.
2. Create your desired genotype
If the phenotype of your mutation varies depending on whether it is heterozygous or homozygous, this can be difficult using the CRISPR-Cas9 system alone. With CRISPR-SNIPER, both DNA strands can be modified easily to obtain your desired genotype – enabling the creation of both homozygous and heterozygous mutations.
3. Increase Screening Specificity
SNIPER increases the specificity of CRISPR-Cas9 by allowing the evaluation of gene editing efficacy under multiple culture conditions. Specificity can be increased further by the addition of nickase, allowing sequences with similarity to your gene of interest to be excluded during the screening process, leaving only the mutants you desire.
4. Track iPSC development
If you are a researcher in developmental biology, you may want to track the differentiation of your iPSCs. Yet reporter genes, such as fluorescence or resistance genes, are often large and therefore difficult to insert. With the enhanced KI rate available with SNIPER, it is possible to insert these functional gene segments more easily – enabling you to track changes in gene expression during differentiation.
5. Edit Multiple Genes Simultaneously
If establishment of your disease model involves multiple modifications, you may need to perform several sequential gene editing experiments. As SNIPER allows rapid optimization of gene editing conditions, this advanced screening technique makes it possible to modify multiple genes simultaneously.
This is achieved by optimizing the culture conditions for a multiplex panel of gRNAs targeted towards your chosen genes, and then using this cocktail to edit your cells in a single experiment. By editing multiple genes at once, you can reduce the negative effects that accompany extended passaging and culture your iPSCs for longer.
Experts in Gene Editing
Did we provide a solution to your gene editing needs today? If not, let us know in the comments. If you are interested in finding out more about REPROCELL’s custom gene editing service, download our CRISPR-SNIPER guide today.
You can also explore our free online resources to learn more about the most advanced gene editing service on the market.
- Meet the creator of SNIPER [WEBINAR]
- Download your free CRISPR-SNIPER Guide [PDF]
- How to determine gene editing success in 2 weeks [BLOG]
†SNIPER is the acronym for Specification of Newly Integrated Position and Exclusion of Random-Integration
*For SNP modification, SNIPER screening detects positive clones at a 30X higher frequency (30%) compared with conventional screening (1%)