While in London I decided to spice up my trip a little. I inquired to the Rihel lab of University College London and accepted an invitation to work with Dr. Sabine Reichert, and Ph.D students Ida and Marcus. In lab I have been doing research on zebra fish sleep/wake cycles, studying behaviors and injecting embryos.
(Left to right) Sabine, Me, Ida, Marcus
Although this trip is an English class on British cultural class, originally joined the class so I could travel. I have never left the country previously, so what better than to go with a bunch of people group and not knowing a single person! If you ask my mother she would tell you I am crazy. All in all, I am having an amazing time and have made life long friends and am currently living in a culture very different from Winona, Minnesota. Cars drive on the left side, using the tube for transportation, and the six hour time difference. (Still has not fully sunk in).
In the Rihel lab, we use zebra fish as model organisms because their circuitry is very similar to ours making them ideal for studying sleep and wake cycles. In the lab, I have been rotating to various projects trying to see as much as I can. So far it has been an amazing experience filled with lots of knowledge. Mean time, I am just trying to soak up as much information that I can.
In a brief summary of what I am doing in lab……
We are trying to create a new genetic line of fish by targeted genome editing with CRISPR/Cas9. CRISPR/Cas9 is used for targeting genome editing to create mutations in the fish. This week we injected the RNA and CRISPR solution into one-stage cell into the zebra fish embryo. The CRISPR will chop up the current double stranded DNA and leave tails so that the RNA will add a different complimentary sequence, ultimately creating a mutation in the genome. We were able to detect a mutation in the DNA with the use of high resolution melt analysis. If the high resolution analysis detects genome editing we will then repeat the injections to test consistency. If there is a mutation again we will then raise up the first generation of fish making it more efficient so injections won’t have to be repeated every time you want a mutated genome. Instead you can set up mating’s and use their embryos for more experiments. In this experiment the goal is to test sleep-wake cycles all with the hopes finding a preventive medicine for insomnia.
With the use of square 96 welled plate, we are able to look at the different movements and behaviors of each fish. A 96 welled plate is where we can put the fish into the wells to monitor each fish’s sleep and wake cycles. Zebra fish are able to soak up applied drugs through their skin making it very effective in targeting all cells. This technique will be helpful in finding what types of drugs prevent and induce sleep.
Dr. Sabine is whom I have mainly been working with in the lab. We have done PCR (polymerase chain reaction) which is used to amplify certain copies of a piece of DNA in a sequence. We have also done a lot of work with in situ hybridization where we insert the correct RNA probe which has a sequence where it can only bind to a specific size and sequence on the DNA strand. By making this reaction occur at a certain temperature, it will make it more favorable for central dogma (DNA—RNA—Protein) to occur. We have also done work with 5-day old embryos by “dissecting the brains”, which is when you remove the skull, jaw, ear, and skin surrounding the brain so the in situ probe can better target the brain. An in situ probe is a technique that allows the sites of expression of particular genes to be detected. The probes target specific mRNA sequences in morphologically preserved tissue or cell preparations by hybridizing the complementary strand of a nucleotide. Dissecting the tissues around the brain it is beneficial so the probe can better reach the desired area in the brain.
Despite all of this research, I have still had time to bond with my classmates. Although they joke…. “Watch out for Mandie, at night she will take your hair samples make zebra fish/human”!
In all reality is completely ridiculous because….
- Cut hair does not contain DNA
- It would be absolutely impossible to make a part zebra fish part human
–Amanda Kaiser
