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CONTACT JAX micetech@jax.org 1.800.422.6423 (US) 1.207.288.5845 (International) LEARN MORE ABOUT JAX MICE AND SERVICES JAX.ORG/JMCRS Frequently Asked Questions What is Bxb1? Dual Bxb1 attP-site "safe harbor landing pad" was inserted in the first intron of Gt(ROSA)26Sor. This enables efficient and precise integration of a donor DNA sequence into the ROSA26 locus by recombinase mediated cassette exchange (RMCE). How large of an insertion can JAX make with Bxb1? We are currently guaranteeing successful insertion of up to 10 kb with Bxb1. If a larger insertion is requested, we will attempt it but will offer it as a fee-for-service project. This technology has been reported to make insertions of up to 40 kb (Low et al, Sci Rep. 2022 Mar 31;12(1). doi: 10.1038/ s41598-022-09445-w.) Can Bxb1 be used for insertions at non-ROSA26 locations? In principle, yes, Bxb1 can be used at nearly any locus. However, the Bxb1 landing pads must first be inserted with CRISPR/Cas9, then Bxb1 can be used to insert the donor template. JAX would consider doing this as a custom multi- stage project upon request. Do Bxb1 projects still use guide RNA(s), similar to CRISPR? No, Bxb1 does not use guide RNA, so these do not need to be designed and validated as seen in CRISPR projects. Will JAX provide a genotyping protocol for the mutation? JAX will provide primer sequences for you to use in your current genotyping platform, as well as sequencing if you would like to design your own primers. Can JAX provide homozygous mice? Yes, we can add on a custom breeding project to breed to homozygosity. We can also breed to other strains (a cre- expressing strain, for example) or produce study-ready cohorts. Do Bxb1 projects still require CRISPR licensing? Yes. CRISPR technology was used to generate the mice used for Bxb1 insertions, therefore CRISPR licensing applies to strains generated using this platform. How do I initiate a new project? Your JAX sales representative will ask for additional details about the model that you would like to generate. Our study directors will then perform a feasibility assessment, and generate a Statement of Work (SOW), which will typically be provided to you within 1 week. If you would like to proceed, you must sign the SOW and provide a PO to initiate the project. Bxb1 Model Generation

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