Knockout of LFNG gene using Di-Cas7-11 system in RAW264.7 macrophages
Faculty Mentor
Jason Ashley
Document Type
Poster
Start Date
10-5-2023 11:15 AM
End Date
10-5-2023 1:00 PM
Location
PUB NCR
Department
Biology
Abstract
Introduction: Cas-711 is a newly discovered CRISPR technology that has been used to make specific incisions within the targeted RNA. Cas-711 prevents collateral damage to the neighboring RNA strands unlike other Cas systems such as Cas-13, which has been known to be destructive to neighboring RNA strands when used for knockout of genes. RAW264.7 macrophages are cells that come from the Abelson leukemia virus which originated in mice and are being used in cellular research. We are looking at the role of the LFNG gene in osteoclast differentiation in RAW264.7 macrophages. These macrophages are used as a model of osteoclast differentiation experiments. Methods: We used Cas-711 to knockout LFNG gene in cultured RAW264.7 macrophages by building a retroviral plasmid and cloning spacer sequences to target mRNA of the LFNG gene. Results: We concluded that the Cas-711 system was able to conduct the knockout of the LFNG gene. We plan on characterizing the differentiation of the RAW264.7 cells.
Recommended Citation
Sanchez, Citlalli, "Knockout of LFNG gene using Di-Cas7-11 system in RAW264.7 macrophages" (2023). 2023 Symposium. 5.
https://dc.ewu.edu/srcw_2023/res_2023/p2_2023/5
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Knockout of LFNG gene using Di-Cas7-11 system in RAW264.7 macrophages
PUB NCR
Introduction: Cas-711 is a newly discovered CRISPR technology that has been used to make specific incisions within the targeted RNA. Cas-711 prevents collateral damage to the neighboring RNA strands unlike other Cas systems such as Cas-13, which has been known to be destructive to neighboring RNA strands when used for knockout of genes. RAW264.7 macrophages are cells that come from the Abelson leukemia virus which originated in mice and are being used in cellular research. We are looking at the role of the LFNG gene in osteoclast differentiation in RAW264.7 macrophages. These macrophages are used as a model of osteoclast differentiation experiments. Methods: We used Cas-711 to knockout LFNG gene in cultured RAW264.7 macrophages by building a retroviral plasmid and cloning spacer sequences to target mRNA of the LFNG gene. Results: We concluded that the Cas-711 system was able to conduct the knockout of the LFNG gene. We plan on characterizing the differentiation of the RAW264.7 cells.
