Two gRNA sequences spanning the TROJAN genomic locus were designed with the CRISPR Design Tool ( A one-step cloning method was used to generate two single gRNAs in LentiCRISPRv2 as previously described (40), with the exception that one of the U6 promoters was replaced with an H1 promoter to avoid potential homologous recombination. The H1 promoter sequence was amplified from pUC-H1-gRNA (Addgene plasmid 61089). The TROJAN deletion efficiency was measured by PCR. The following gRNA was used for ZMYND8 knockout: 5′-GTGATGTGTCCTGCGGCGAG-3′. Single clones were established by dilution cloning.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.