What is CRISPR/Cas9?
CRISPR or clustered regularly interspaced short palindromic repeats was first discovered in 1987 when Dr. Ishino and colleagues noticed a strange repetitive DNA sequence in the bacteria Escherichia coli. Following studies observed a similar sequence pattern in other bacteria and archaea and later came to understand that in-between these repeats there were DNA sequences that corresponded to bacteriophages that had previously infected the bacteria or archaea! That’s right, the CRISPR/Cas family of DNA sequences are bacteria and archaea’s natural immune system.
What essentially happens is when bacteria and archaea are invaded by bacteriophage, they target the bacteriophages DNA and send an enzyme (Cas Protein) with this information to cut up the phages DNA, killing the phage and surviving to live another day.
In 2012, researchers utilized this natural immune system and made slight changes so that they could target any gene that they were interested in, using Streptococcus pyogenes Cas9 enzyme. Currently, CRISPR/Cas9 has become a household name as it is an easy to use genome editing system and is being used to fight cancer, edit crops, blindness, and study the many genes and organisms that we currently do not understand well.
Want to learn more? We recommend watching this awesome video from the the McGovern Institute at MIT
&/or listening to these Radiolab Podcasts:
How are we trying to use CRISPR/Cas9 with Dinoflagellates?
If you guys remember from our Dinoflagellate Gene Transformation page, dinoflagellates are SUPER important for the environment but we also don’t understand them very well. This is because more than 50% of their genes are novel and they are very different from most other known eukaryotes. We aim to use CRISPR/Cas9 to study these unique genes to try to understand dinoflagellates even better.
We have had success using CRISPR/Cas9 with the diatom Phaeodactylum tricornutum and are leveraging our experience in this model organism to design protocols for the dinoflagellate Oxyrrhis marina.
Funded by the Gordon and Betty Moore Foundation-Marine Microbiology Initiative (MMI), the over goal of this endeavor is to develop gene editing in dinoflagellates. This include two major objectives:
To transform dual gene cassettes and optimize our gene transformation efforts.
Develop a protocol for CRISPR-Cas9 for dinoflagellates.
Our protocol for CRISPR-Cas9 gene editing in Phaeodactylum tricornutum: https://www.protocols.io/view/introducing-dinoflagellate-gene-into-diatom-using-hm2b48e
This is still preliminary work for dinoflagellates but we invite you to look at our on-going protocol: https://www.protocols.io/view/protocol-for-introducing-fluorescently-labeled-cri-bei2jcge