Genetic Manipulation of B Cell Mice

Mice used in this study were group housed (up to 5 mice of the same sex) with unrestricted access to water and standard chow diet, unless otherwise indicated, under specific pathogen free conditions in the Rockefeller University (RU) Comparative Bioscience Center. Mice used in this study ranged from 6 to 14 weeks old. Wild-type C57BL/6J and B6.SJL male mice were purchased from Jackson Laboratories. B1-8^hi, B1-8^hi DEC205^−/−, B1-8^hi CFP, and B1-8^hi c-Myc-GFP have been described^{6 (link),20 (link),75 (link)}. BTK^C481S point mutation mice were generated by microinjection of gRNA, hCas9, and single-stranded donor oligonucleotides into B6 zygote pronuclei (RU CRISPR and Genome Editing Center, RU Transgenic and Reproductive Technology Center)^{76 (link)}. Mutants were backcrossed to B6.SJL for 5+ generations to remove possible CRISPR off-target effects. B1-8^hi BTK^C481S and B1-8^hi DEC205^−/− BTK^C481S were generated by crossing to B1-8^hi and B1-8^hi DEC205^−/−. All experiments conform to protocols approved by the RU Institutional Animal Care and Use Committee.

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Chen S.T., Oliveira T.Y., Gazumyan A., Cipolla M, & Nussenzweig M.C. (2023). B cell receptor signaling in germinal centers prolongs survival and primes B cells for selection. Immunity, 56(3), 547-561.e7.

Publication 2023

B6.SJL male mice

Crispr Diet Donor Institutional animal care and use committee Male Mice Microinjection Oligonucleotides Point mutation Reproductive technology Specific pathogen free Transgenic Zygote

Corresponding Organization : Howard Hughes Medical Institute

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Variable analysis

independent variables

Genotype: Wild-type C57BL/6J, B1-8hi, B1-8hi DEC205-/-, B1-8hi CFP, B1-8hi c-Myc-GFP, BTKbC481S, B1-8hi BTKbC481S, B1-8hi DEC205-/- BTKbC481S

dependent variables

Not explicitly mentioned

control variables

Mouse age: 6 to 14 weeks old
Housing: Group housed (up to 5 mice of the same sex) with unrestricted access to water and standard chow diet, unless otherwise indicated, under specific pathogen free conditions
Strain: B6.SJL mice used for backcrossing BTKbC481S mutants

controls

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

Annotations

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