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Immunization experiment scheme for in vivo screen Host mice (IOMA gl) were immunized with OVA in alum. At week 2 after immunization, B cells from B1-8 hi , <t>LSL-Cas9,</t> AID-Cre, Kappa KO animals were transduced with pooled sgRNA library retrovirus and adoptively transferred to the hosts. Subsequently, the host animals were boosted with NP-OVA in alum at day 0 and 2 and administered with anti-DEC205-OVA mAb at day 6.5. Spleens were harvested and processed for cell sorting at day 10.
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Immunization experiment scheme for in vivo screen Host mice (IOMA gl) were immunized with OVA in alum. At week 2 after immunization, B cells from B1-8 hi , <t>LSL-Cas9,</t> AID-Cre, Kappa KO animals were transduced with pooled sgRNA library retrovirus and adoptively transferred to the hosts. Subsequently, the host animals were boosted with NP-OVA in alum at day 0 and 2 and administered with anti-DEC205-OVA mAb at day 6.5. Spleens were harvested and processed for cell sorting at day 10.
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Immunization experiment scheme for in vivo screen Host mice (IOMA gl) were immunized with OVA in alum. At week 2 after immunization, B cells from B1-8 hi , <t>LSL-Cas9,</t> AID-Cre, Kappa KO animals were transduced with pooled sgRNA library retrovirus and adoptively transferred to the hosts. Subsequently, the host animals were boosted with NP-OVA in alum at day 0 and 2 and administered with anti-DEC205-OVA mAb at day 6.5. Spleens were harvested and processed for cell sorting at day 10.
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Immunization experiment scheme for in vivo screen Host mice (IOMA gl) were immunized with OVA in alum. At week 2 after immunization, B cells from B1-8 hi , <t>LSL-Cas9,</t> AID-Cre, Kappa KO animals were transduced with pooled sgRNA library retrovirus and adoptively transferred to the hosts. Subsequently, the host animals were boosted with NP-OVA in alum at day 0 and 2 and administered with anti-DEC205-OVA mAb at day 6.5. Spleens were harvested and processed for cell sorting at day 10.
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Immunization experiment scheme for in vivo screen Host mice (IOMA gl) were immunized with OVA in alum. At week 2 after immunization, B cells from B1-8 hi , <t>LSL-Cas9,</t> AID-Cre, Kappa KO animals were transduced with pooled sgRNA library retrovirus and adoptively transferred to the hosts. Subsequently, the host animals were boosted with NP-OVA in alum at day 0 and 2 and administered with anti-DEC205-OVA mAb at day 6.5. Spleens were harvested and processed for cell sorting at day 10.
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Immunization experiment scheme for in vivo screen Host mice (IOMA gl) were immunized with OVA in alum. At week 2 after immunization, B cells from B1-8 hi , <t>LSL-Cas9,</t> AID-Cre, Kappa KO animals were transduced with pooled sgRNA library retrovirus and adoptively transferred to the hosts. Subsequently, the host animals were boosted with NP-OVA in alum at day 0 and 2 and administered with anti-DEC205-OVA mAb at day 6.5. Spleens were harvested and processed for cell sorting at day 10.
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Immunization experiment scheme for in vivo screen Host mice (IOMA gl) were immunized with OVA in alum. At week 2 after immunization, B cells from B1-8 hi , LSL-Cas9, AID-Cre, Kappa KO animals were transduced with pooled sgRNA library retrovirus and adoptively transferred to the hosts. Subsequently, the host animals were boosted with NP-OVA in alum at day 0 and 2 and administered with anti-DEC205-OVA mAb at day 6.5. Spleens were harvested and processed for cell sorting at day 10.

Journal: STAR Protocols

Article Title: Protocol for an in vivo CRISPR screen for germinal center B cells in mice using ecotropic retrovirus

doi: 10.1016/j.xpro.2026.104586

Figure Lengend Snippet: Immunization experiment scheme for in vivo screen Host mice (IOMA gl) were immunized with OVA in alum. At week 2 after immunization, B cells from B1-8 hi , LSL-Cas9, AID-Cre, Kappa KO animals were transduced with pooled sgRNA library retrovirus and adoptively transferred to the hosts. Subsequently, the host animals were boosted with NP-OVA in alum at day 0 and 2 and administered with anti-DEC205-OVA mAb at day 6.5. Spleens were harvested and processed for cell sorting at day 10.

Article Snippet: Mouse: 6-weeks-old, sex-matched LSL-Cas9: B6J.129(B6N)-Gt(ROSA)26Sortm1(CAG-cas9∗,-EGFP)Fezh/J , The Jackson Laboratory , cat# 026175.

Techniques: In Vivo, Transduction, FACS