Grant applications proposing work that will be performed by Cellular Engineering (CE) or with CE’s assistance do not require letters of support unless the methods that will be used are not included in CE’s service description in the JAX Facilities and Other Resources master document (see description below). For methods that are unusual or particularly challenging in the iPSC field, or that were recently published, please contact the CE director (CellularEngineeringSVC@jax.org) whether a letter of support would be appropriate.
Facilities and Other Resources Documentation:
The Cellular Engineering laboratory (CE) provides high-quality, custom-edited lines of human induced pluripotent stem cells (iPSCs) using CRISPR/Cas technologies, offering a range of allele types that include knockouts, knockins, single-base variants, and conditional loss-of-function alleles. Precision editing is based on high-efficiency co-delivery of Cas9 or Cas12a RNP and repair templates (single-stranded or plasmid donor DNA) using the Amaxa™ 4D-Nucleofector™ System (Lonza). The CE cell culture space has six NuAire Class II, Type A2 biological safety cabinets, including one set up with an Olympus SZ61 stereo microscope with interlock dual-light guide illumination used for isolation of edited iPSC clones. Automated brightfield and fluorescent cell counting is performed with a LUNA-FX7 counter. PCR-based genotyping of edited clones is performed in the CE molecular biology lab, which houses five C1000 Touch™ Thermal Cyclers (Bio-Rad), a DS-11 Fx+ spectrophotometer (DeNovix), and a NuGenius gel documentation workstation (Syngene). A QX200 AutoDG Droplet Digital PCR system (Bio-Rad) is housed in the CE molecular lab and is available to trained users for digital droplet (ddPCR). CE works closely with the Genome Technologies and Flow Cytometry services at JAX. Extensive quality control of clonal lines is available to confirm the molecular structure of the engineered alleles, and to identify potential off-target damage at highly related sites and other genomic rearrangements that may occur during culture. Additional equipment includes a HemaVet® 950 FS Hematology System (Drew Scientific) for enumerating cell populations in blood. Lastly, CE provides consultation on allele design, guide RNA selection, and editing protocols.
Cellular Engineering equipment
Acknowledgement & Authorship: (sample language for Scientific Services acknowledgements for your team will be included here)
Cellular Engineering is a collaborative core facility with specialized expertise. Please consider the importance of our contributions to your research program which may fit one or several roles of the CRediT authorship taxonomy.
Please remember to acknowledge the use of the JAX Cellular Engineering service in manuscripts and grant applications:
“We gratefully acknowledge the Cellular Engineering service at The Jackson Laboratory for [examples: 1) expert assistance with XXXX; or 2) generating the XXXX knockout iPSC line].”
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