CRISPR-Mediated Triple Knockout of SLAMF1, SLAMF5 and SLAMF6 Supports Positive Signaling Roles in NKT Cell Development.
The SLAM family receptors contribute to diverse aspects of lymphocyte biology and signal via the small adaptor molecule SAP. Mutations affecting SAP lead to X-linked lymphoproliferative syndrome Type 1, a severe immunodysregulation characterized by fulminant mononucleosis, dysgammaglobulinemia, and lymphoproliferation/lymphomas. Patients and mice having mutations affecting SAP also lack germinal centers due to a defect in T:B cell interactions and are devoid of invariant NKT (iNKT) cells. However, which and how SLAM family members contribute to these phenotypes remains uncertain. Three SLAM family members: SLAMF1, SLAMF5 and SLAMF6, are highly expressed on T follicular helper cells and germinal center B cells. SLAMF1 and SLAMF6 are also implicated in iNKT development. Although individual receptor knockout mice have limited iNKT and germinal center phenotypes compared to SAP knockout mice, the generation of multi-receptor knockout mice has been challenging, due to the genomic linkage of the genes encoding SLAM family members. Here, we used Cas9/CRISPR-based mutagenesis to generate mutations simultaneously in Slamf1, Slamf5 and Slamf6. Genetic disruption of all three receptors in triple-knockout mice (TKO) did not grossly affect conventional T or B cell development and led to mild defects in germinal center formation post-immunization. However, the TKO worsened defects in iNKT cells development seen in SLAMF6 single gene-targeted mice, supporting data on positive signaling and potential redundancy between these receptors.
Genetic Disease Research Branch, National Human Genome Research Institute, Bethesda, Maryland, United States of America.
Bonnie
B
Huang
Genetic Disease Research Branch, National Human Genome Research Institute, Bethesda, Maryland, United States of America.
Julio
J
Gomez-Rodriguez
Genetic Disease Research Branch, National Human Genome Research Institute, Bethesda, Maryland, United States of America.
Silvia
S
Preite
Embryonic Stem Cell and Transgenic Mouse Core, National Human Genome Research Institute, Bethesda, Maryland, United States of America.
Lisa J
LJ
Garrett
Genomics Core, National Human Genome Research Institute, National Human Genome Research Institute, Bethesda, Maryland, United States of America.
Ursula L
UL
Harper
Genetic Disease Research Branch, National Human Genome Research Institute, Bethesda, Maryland, United States of America.
Pamela L
PL
Schwartzberg
D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D018858:Germinal Center / Q000378:metabolism*; D008213:Lymphocyte Activation / Q000502:physiology; D051379:Mice; D018345:Mice, Knockout; D055611:Natural Killer T-Cells / Q000378:metabolism*; D015398:Signal Transduction; D000071176:Signaling Lymphocytic Activation Molecule Family / Q000235:genetics* / Q000378:metabolism; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1 / Q000235:genetics* / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism
C498971:Cd84 protein, mouse; D000071176:Signaling Lymphocytic Activation Molecule Family; C000604063:Slamf6 protein, mouse; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1