Extended Data Fig. 10: Method for rapid cloning of augmented sgRNAs into TRV vectors.
From: Multiplexed heritable gene editing using RNA viruses and mobile single guide RNAs
A cloning vector was developed that allows for simple and efficient cloning of unique sgRNA target sequences into TRV2 vectors. Unique sgRNAs are added to the vector using one unique primer (that contains the sgRNA target of interest) and one reverse primer (that can be used for all unique primers). These primers are used to amplify the modified sgRNA from the TRV2 cloning vector backbone. This amplicon is then added to the TRV2 cloning vector through a Golden Gate Assembly cloning step to create a final TRV2 vector with the modified sgRNA of interest.
