Ralitsa Madsen is a PhD student in the University of Cambridge. Below she shares her journey in harnessing the power of CRISPR. Ultimately, she was able to create point mutation knockins in HEK293s using CRISPR with up to 18% efficiency without selection. Check out this post for her tips and tricks!
When I started my PhD project in October 2015, I knew three things for sure:
- My project involved modelling disease-specific point mutations in human pluripotent stem cells.
- I would generate these point mutations using CRISPR/Cas9, a technology providing scientists with state-of-the-art molecular DNA scissors.
- I had never used CRISPR/Cas9 and did not have a clue what conditions would work best in my hands!
It is now April 2016, and I have come a long way. Following multiple rounds of optimisations, trials-and-errors and long working hours, I have become a confident user of this technology. However, this progress would have been impossible had it not been for the multitude of protocols that have been published across scientific journals and blogs around the web. It is time for another blog post of this kind, now with me in the writer’s seat, taking you through my CRISPR journey.
To read the full post, please follow this link: https://blog.benchling.com/no-more-cloning-quick-and-easy-crispr-with-sgrna-gene-fragments/