Generating CS1-Deficient MM.1S Multiple Myeloma Cells

MM.1S cells, kindly provided by Leif Bergsagel, were modified to MM.1S-CG (16 (link)). ΔCS1-MM.1S-CGs were generated using the CS1 guide sequence above cloned into pMLM3636 (Addgene) lentiviral vector. MM1.S-CG cells were transduced with 1ug gRNA plasmid, 250ng Cas9-HF plasmid and electroporated in SF solution using the Lonza 4D Nucleofector DS-137 program. CS1 negative cells were sorted using a MoFlow. OPM2 cells (DMSZ, Germany) were modified to express CBR-GFP as above.

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O’Neal J., Ritchey J.K., Cooper M.L., Niswonger J., Sofía González L., Street E., Rettig M.P., Gladney S.W., Gehrs L., Abboud R., Prior J.L., Haas G.J., Jayasinghe R.G., Ding L., Ghobadi A., Vij R, & DiPersio J.F. (2022). CS1 CAR-T targeting the distal domain of CS1 (SLAMF7) shows efficacy in high tumor burden myeloma model despite fratricide of CD8+CS1 expressing CAR-T cells. Leukemia, 36(6), 1625-1634.

Publication 2022

pMLM3636 4D Nucleofector DS-137

Cells Plasmid Vector

Corresponding Organization : Washington University in St. Louis

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Variable analysis

independent variables

Modification of MM.1S cells to MM.1S-CG
Generation of ΔCS1-MM.1S-CGs using the CS1 guide sequence cloned into pMLM3636 lentiviral vector
Transduction of MM1.S-CG cells with 1ug gRNA plasmid and 250ng Cas9-HF plasmid
Modification of OPM2 cells to express CBR-GFP

dependent variables

CS1 negative cells sorted using a MoFlow

control variables

Cells were electroporated in SF solution using the Lonza 4D Nucleofector DS-137 program

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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