I am new to cloning and have recently carried out some cloning using the Gateway system with a lot of help from some really helpful members in this forum!
I have to carry out some gene knockouts now and although I am familiar with Crispr/Cas9 knockouts, I have no knowledge of how to design guides and clone them into appropriate vectors. I had a number of questions I was hoping I could get some help on.
1. I am aware that there are numerous methods of delivering the Crispr/Cas9 system into cells but not sure what all he different methods are? Can someone explain to me the various methods. My current lab uses the Zhang lab method with the lentiviral delivery system but I would like to find out about what other vectors/systems people use?
2. Can someone help me design guides for my gene of interest/give me step by step instructions on how to do this?
3. How do I incorporate the guide into my lentiviral vector?
Any help would be greatly appreciated!