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GEO help: Mouse over screen elements for information. |
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Status |
Public on Jan 03, 2025 |
Title |
Advancing the genetic engineering toolbox by combining AsCas12a knock-in mice with ultra-compact screening |
Organism |
Mus musculus |
Experiment type |
Other
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Summary |
Cas12a is a next-generation gene-editing tool that enables multiplexed gene targeting. Here, we present a mouse model that constitutively expresses enhanced Acidaminococcus sp. Cas12a (enAsCas12a) linked to an mCherry fluorescent reporter. We demonstrate efficient single and multiplexed gene-editing in vitro, using primary and transformed cells from enAsCas12a mice. We further demonstrate successful in vivo gene-editing, using normal and cancer-prone enAsCas12a stem cells to reconstitute the haematopoietic system of wild-type mice. We also present compact, genome-wide Cas12a knockout libraries, with four crRNAs per gene encoded across one (Scherzo) or two (Menuetto) vectors, and demonstrate the utility of these libraries across methodologies: in vitro enrichment and drop-out screening in lymphoma cells and immortalised fibroblasts, respectively, and in vivo screens to identify lymphoma-driving events. Finally, we demonstrate CRISPR multiplexing via simultaneous gene knockout (via Cas12a) and activation (via dCas9-SAM) using primary T cells and fibroblasts. Our enAsCas12a mouse and accompanying crRNA libraries enhance genome engineering capabilities and complement current CRISPR technologies.
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Overall design |
NGS of mouse cells containing CRISPR/Cas12 libraries (Menuetto/dual and Scherzo/quad). Sequences are of the sgRNA plasmids. Three screens were performed: 1) enrichment screen in mouse EuMyc lymphoma cells using S63845 and Nutlin-3a as selection pressures, 2) drop out screen in immortalised mouse dermal fibroblasts (MDFs), and 3) in vivo enrichment screen for drivers of lymphomagenesis in mice.
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Contributor(s) |
La Marca JE, Diepstraten ST, Deng Y, Jin W, Herold M, Fortin J, Haley B |
Citation missing |
Has this study been published? Please login to update or notify GEO. |
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Submission date |
Jan 03, 2025 |
Last update date |
Jan 03, 2025 |
Contact name |
John E La Marca |
Organization name |
Walter and Eliza Hall Institute
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Department |
Blood Cells and Blood Cancer
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Street address |
1G Royal Parade
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City |
Parkville |
State/province |
VIC |
ZIP/Postal code |
3052 |
Country |
Australia |
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Platforms (2) |
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Samples (86)
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GSM8708884 |
EuMyc cells, Menuetto, Input, Rep4 |
GSM8708885 |
EuMyc cells, Menuetto, Input, Rep5 |
GSM8708886 |
EuMyc cells, Menuetto, Input, Rep6 |
GSM8708887 |
EuMyc cells, Menuetto, DMSO, Rep1 |
GSM8708888 |
EuMyc cells, Menuetto, DMSO, Rep2 |
GSM8708889 |
EuMyc cells, Menuetto, DMSO, Rep3 |
GSM8708890 |
EuMyc cells, Menuetto, DMSO, Rep4 |
GSM8708891 |
EuMyc cells, Menuetto, DMSO, Rep5 |
GSM8708892 |
EuMyc cells, Menuetto, DMSO, Rep6 |
GSM8708893 |
EuMyc cells, Menuetto, Nutlin, Rep1 |
GSM8708894 |
EuMyc cells, Menuetto, Nutlin, Rep2 |
GSM8708895 |
EuMyc cells, Menuetto, Nutlin, Rep3 |
GSM8708896 |
EuMyc cells, Menuetto, Nutlin, Rep4 |
GSM8708897 |
EuMyc cells, Menuetto, Nutlin, Rep5 |
GSM8708898 |
EuMyc cells, Menuetto, Nutlin, Rep6 |
GSM8708899 |
EuMyc cells, Menuetto, S63845, Rep1 |
GSM8708900 |
EuMyc cells, Menuetto, S63845, Rep2 |
GSM8708901 |
EuMyc cells, Menuetto, S63845, Rep3 |
GSM8708902 |
EuMyc cells, Menuetto, S63845, Rep4 |
GSM8708903 |
EuMyc cells, Menuetto, S63845, Rep5 |
GSM8708904 |
EuMyc cells, Menuetto, S63845, Rep6 |
GSM8708905 |
EuMyc cells, Scherzo, Input, Rep1 |
GSM8708906 |
EuMyc cells, Scherzo, Input, Rep2 |
GSM8708907 |
EuMyc cells, Scherzo, Input, Rep3 |
GSM8708908 |
EuMyc cells, Scherzo, Input, Rep4 |
GSM8708909 |
EuMyc cells, Scherzo, Input, Rep5 |
GSM8708910 |
EuMyc cells, Scherzo, Input, Rep6 |
GSM8708911 |
EuMyc cells, Scherzo, DMSO, Rep1 |
GSM8708912 |
EuMyc cells, Scherzo, DMSO, Rep2 |
GSM8708913 |
EuMyc cells, Scherzo, DMSO, Rep3 |
GSM8708914 |
EuMyc cells, Scherzo, DMSO, Rep4 |
GSM8708915 |
EuMyc cells, Scherzo, DMSO, Rep5 |
GSM8708916 |
EuMyc cells, Scherzo, DMSO, Rep6 |
GSM8708917 |
EuMyc cells, Scherzo, Nutlin, Rep1 |
GSM8708918 |
EuMyc cells, Scherzo, Nutlin, Rep2 |
GSM8708919 |
EuMyc cells, Scherzo, Nutlin, Rep3 |
GSM8708920 |
EuMyc cells, Scherzo, Nutlin, Rep4 |
GSM8708921 |
EuMyc cells, Scherzo, Nutlin, Rep5 |
GSM8708922 |
EuMyc cells, Scherzo, Nutlin, Rep6 |
GSM8708923 |
EuMyc cells, Scherzo, S63845, Rep1 |
GSM8708924 |
EuMyc cells, Scherzo, S63845, Rep2 |
GSM8708925 |
EuMyc cells, Scherzo, S63845, Rep3 |
GSM8708926 |
EuMyc cells, Scherzo, S63845, Rep4 |
GSM8708927 |
EuMyc cells, Scherzo, S63845, Rep5 |
GSM8708928 |
EuMyc cells, Scherzo, S63845, Rep6 |
GSM8708929 |
iMDF cells, Menuetto, T0, Rep1 |
GSM8708930 |
iMDF cells, Menuetto, T0, Rep2 |
GSM8708931 |
iMDF cells, Menuetto, T0, Rep3 |
GSM8708932 |
iMDF cells, Menuetto, T1, Rep1 |
GSM8708933 |
iMDF cells, Menuetto, T1, Rep2 |
GSM8708934 |
iMDF cells, Menuetto, T1, Rep3 |
GSM8708935 |
iMDF cells, Menuetto, T2, Rep1 |
GSM8708936 |
iMDF cells, Menuetto, T2, Rep2 |
GSM8708937 |
iMDF cells, Menuetto, T2, Rep3 |
GSM8708938 |
iMDF cells, Menuetto, T3, Rep1 |
GSM8708939 |
iMDF cells, Menuetto, T3, Rep2 |
GSM8708940 |
iMDF cells, Menuetto, T3, Rep3 |
GSM8708941 |
Tumour, Menuetto, Mouse1124 |
GSM8708942 |
Tumour, Menuetto, Mouse1125 |
GSM8708943 |
Tumour, Menuetto, Mouse1126 |
GSM8708944 |
Tumour, Menuetto, Mouse1127 |
GSM8708945 |
Tumour, Menuetto, Mouse1128 |
GSM8708946 |
Tumour, Menuetto, Mouse1130 |
GSM8708947 |
Tumour, Menuetto, Mouse1131 |
GSM8708948 |
Tumour, Menuetto, Mouse1141 |
GSM8708949 |
Tumour, Menuetto, Mouse1142 |
GSM8708950 |
Tumour, Menuetto, Mouse1143 |
GSM8708951 |
Tumour, Menuetto, Mouse1144 |
GSM8708952 |
Tumour, Menuetto, Mouse1145 |
GSM8708953 |
Tumour, Menuetto, Mouse1146 |
GSM8708954 |
Tumour, Menuetto, Mouse1147 |
GSM8708955 |
Tumour, Menuetto, Mouse1148 |
GSM8708956 |
Tumour, Menuetto, Mouse1150 |
GSM8708957 |
Tumour, Menuetto, Mouse1151 |
GSM8708958 |
Cell line, Menuetto, Mouse 1126 |
GSM8708959 |
Cell line, Menuetto, Mouse 1128 |
GSM8708960 |
Cell line, Menuetto, Mouse 1130 |
GSM8708961 |
Cell line, Menuetto, Mouse 1131 |
GSM8708962 |
Cell line, Menuetto, Mouse 1141 |
GSM8708963 |
Cell line, Menuetto, Mouse 1142 |
GSM8708964 |
Cell line, Menuetto, Mouse 1145 |
GSM8708965 |
Cell line, Menuetto, Mouse 1146 |
GSM8708966 |
Cell line, Menuetto, Mouse 1147 |
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Relations |
BioProject |
PRJNA1206072 |
Supplementary file |
Size |
Download |
File type/resource |
GSE285778_EuMycCountMenuetto.txt.gz |
1.0 Mb |
(ftp)(http) |
TXT |
GSE285778_EuMycCountScherzo.txt.gz |
624.3 Kb |
(ftp)(http) |
TXT |
GSE285778_InVivoCount.txt.gz |
267.1 Kb |
(ftp)(http) |
TXT |
GSE285778_iMDFCount.txt.gz |
1.4 Mb |
(ftp)(http) |
TXT |
SRA Run Selector |
Raw data are available in SRA |
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