Today, marked yet another major step towards our goal. Dr. Jana Marcette requested that our team in St. Louis, MO get together and meet to discuss our plan of action as what has seemingly been a dream is quickly becoming a reality! So, Dr. Marcette, Dr. David Curiel, and myself met in Dr. Curiel’s office (12/14/18) to discuss research plans moving forward. We are going to be developing a database that will include the contact information of scientists interested in CAMSAP1 so that we can all readily collaborate. Most importantly, we were given another update by The Jackson Laboratory (Jax)!
To my surprise, upon entering Dr. Curiel’s office, I was handed a packet sent by Jax that contained news that another WONDERFUL research milestone had been reached and completed! Of the 45 pups created from the microinjection process, FOUR of them were identified following genetic screening as “Founder Mice” – the mice that had successfully been genetically created to be carriers of the gene mutation, yet able to transmit the mutation to future generations, but who remain outwardly/phenotypically unaffected.
These CRISPR/Cas9-generated mice were transferred to the Jax mating facility to be bred with “wild-type” mice to create second-generation (N1) offspring. By using wild-type mice, it assures that germline transmission can be established – meaning that future generations may be able to also pass along the CAMSAP1 gene mutation. If the laws of genetics hold true, roughly 50% of the offspring from this mating process will produce effected homozygous CAMSAP1-deficient mice that will be analyzed/phenotyped to understand the effect of any mutations created. I’ve borrowed this image to depict how this process takes place – the progress is currently at the level of the first two mice below (image hyperlinked to source):
Here is an idea of the timeline – we are now at the “Founder report packet” step (per The Jackson Laboratory CAMSAP1 Knockout with CRISPR/Cas9 “Founder Report”):
As mentioned in our November Research Update, the birth of the Founder Mice (N0) occurred on 11/03/2018, our wedding anniversary. If the time line above holds true, even more irony may take place in the near future:
The “Landon-like” N1 pups could potentially be born on 01/04/2018, which is Landon’s birthday…
More updates to come in a few short weeks so please stay tuned. Happy holidays and here is to a promising and successful 2019!
An INCREDIBLE day! A second major research hurdle was crossed yesterday. We received this email from The Jackson Laboratory:
“[We] just got word today that the microinjection for the Camsap1 KO project has successfully produced 45 pups born on 11.3.18. These animals will be weaned and sampled at 3 weeks of age, and then the DNA sequence will be analyzed to identify potential founders mice.
I hope this is good news for you!”
So…what does this mean? First, THIS IS ABSOLUTELY GREAT NEWS!!! The irony? This first generation of mice (called N0) were all born on 11.3.2018, which happens to be Lauren and I’s wedding anniversary!
Simply, the 45 first generation mice will be screened at 3 weeks old to identify the ones that are carriers of CAMSAP1 but unaffected like Lauren and I. Then, the lab will breed those mice. Approximately 25% of their offspring (the second generation, or N1) should have Landon’s exact CAMSAP1 mutation. These N1 mice with Landon’s mutation will be the ones that come to St. Louis to undergo gene therapy/rescue by Dr. Curiel.
Scientifically, to simplify (as much as possible), 45 mice were born following a procedure whereby a microscopic needle was inserted into a developing mouse zygote following initial fertilization. The needle was used to inject the developing cell with the necessary genetic CRISPR/CAS9 enzyme (think of it like a scissors) and guide arm (to guide the scissors to the exact gene) to knockout (KO) or remove the CAMSAP1 gene at the site of Landon’s mutation (see image below linked to and borrowed from The Jackson Laboratory website).
The wait is over: We are thrilled to share that we have our first research progress update! 🎉
Before the mouse model research can begin onsite at Washington University in St. Louis School of Medicine, a laboratory in Maine called The Jackson Laboratory is working towards creating mice that carry Landon’s gene mutation. The process of creating Landon’s mice begins at a cellular level and takes months. Please understand that the world of medical research can seem very slow-moving, yet can be very precise and successful. Thus, we must remind ourselves to be patient.
The Jackson Laboratory has acquired the necessary RNA (to simplify, RNA is like one side of a zipper on a coat – two RNAs equals DNA) that will direct a specific enzyme called Cas9 to “cut” the CAMSAP1 gene in the exact location where Landon’s gene has a mutation. This creates a modified gene. The modified gene was then reintroduced to normal mouse cells. These modified mouse cells have been tested and found to have no traces of the CAMSAP1 gene, which mimics Landon’s cells! SUCCESS!!! This will not only help Landon, but it will also help the nearly 500 people worldwide who’ve been identified as having the same mutated CAMSAP1 gene.
The next step is to introduce this exact RNA and Cas9 enzyme into hundreds of mouse zygotes. The zygote is the cell formed when the egg and sperm unite to make a single cell, which will divide and grow up to be a mouse. If this is also successful, mice will be born about 3 weeks later carrying Landon’s mutation!
09/15/2018: A Night at the Mouse Races Event
06/26/2018: Payment delivered to Washington University to purchase a mouse model of CAMSAP1 genetic mutation.
06/17/2018: First fundraiser: T-Shirt campaign kickoff
06/13/2018: Landon’s League Foundation founded as 501(c)3 nonprofit organization.
06/13/2018: Initial quote for a mouse model of CAMSAP1 deficiency received from Jackson Labs.
5/23/2018: Meeting with genetic therapist at Washington University in St. Louis who agreed to research CAMSAP1 in an effort to create a gene therapy for individuals effected by a defect in CAMSAP1.