CRISPR, also known as Clustered Regularly Interspaced Short Palindromic Repeats, is a specific component of the bacterial immune system that can penetrate through the DNA. For the past couple of years, CRISPR has been considered a gene editing tool due to its effectiveness. CRISPR works like a scissor to cut down the DNA molecules through the customizable guide.
The CRISPR system is made of two different parts such as one Cas nuclease which is capable of cutting and binding the DNA and a gRNA sequence that helps the Cas nuclease reach the target.
CRISPR is extremely important as it helps scientists rewrite different types of genetic codes in different types of organisms. CRISPR is more effective, cheaper, and simpler than the previous gene editing options. Let’s discuss how you can use CRISPR effectively.
Guide the Design of the RNA
The guide RNA design is one of the most important aspects you need to keep in mind if you want to design a successful CRISPR experiment. The sgRNA you choose will depend on numerous things such as the type of editing you need, whether it’s KI, KO, or prime editing. As per Genetic Education, sgRNA are the sequences of RNA molecules. The guide RNA design will also play a massive role in the CRISPRa and CRISPRi processes.
It doesn’t matter which CRISPR design tool you use, you need to make sure you know the correct process of using the tool so that you don’t end up making mistakes. You also need to pay close attention to the requirements of your experiment while choosing a CRISPR gene editing tool.
Choose the Perfect Cas9 Variant
The Cas nuclease you choose will also determine the overall success of your experiment. This is because Cas9 was not only the first but also the foremost substance used in the Cas nuclease, where it’s originally isolated from the SpCas9 or bacterial strain Streptococcus pyogenes. However, keep in mind that there are numerous naturally occurring as well as engineered variants of Cass available that will prove effective in different situations. This use of CRISPR technology will prove beneficial.
For instance, engineered Cas9 can be used by scientists who have plans to create single-stranded breaks as well as different types of high-quality Cas9 variants to reduce off-target edits. On the other hand, naturally occurring Cas 13 will be used for diagnostic potential.
This is another important thing you need to know. There is no specific or universal transfection protocol for the experiments. This is because the delivery process of editing the different components of genes is depending on the organisms as well as cells you’re targeting. Not to mention, it will also depend on the delivery of the type of editing cargo. There are different transfection protocols available in the market and you need to choose something as per your needs and requirements. Make sure you don’t make mistakes while choosing transfection protocols.
When you choose the perfect transfection protocols, you will be able to analyze your CRISPR experiment without any problems. But it needs to be tailored as per your experiment.
This is the ultimate guide you need to know about CRISPR. Consider letting us know if you have any queries or comments howitstart.