Per Winge - CRISPR/Cas in eukaryotic cells and organisms Flashcards
Inactivation of genes has been an important tool to study the functions of genes in many model organisms. What is the advantage of the CRISPR/Cas9 technology compared to the previous knockout methods?
Bacteria have CRISPR/Cas systems that often express a trans activating CRISPR RNA (trcrRNA), what is the function of the trcrRNA?
The function of the trcRNA is to facilitate DNA cutting by the Cas protein.
The Cas9 protein contain two nuclease domains, the RuvC and HNH domains. If the catalytic sites in RuvC and HNH are inactivated the protein cannot produce double stranded DNA breaks and is called a dead-Cas9 (dCas9). Can the dCas9 still be of any use? Describe two examples of how it can be used.
Bacteria uses CRISPR/Cas systems to protect themselves from invading bacteriophages. This has led to an arms race where the bacteriophages try to
evade the “CRISPR defence”. In this process some bacteriophages express anti-CRISPR proteins that inhibits Cas9. Can anti-CRISPR proteins have any
useful applications in biotechnology?
How the CRISPR/Cas system are delivered to eukaryotic cells may vary from organism to organism. What are the challenges related to this delivery process? How do we produce or engineer cells that contain Cas proteins and guide RNAs?
