Modular safe-harbor transgene insertion (MosTI) for targeted single-copy and extrachromosomal array integration in C. elegans.
El Mouridi S, Alkhaldi F, Frokjaer-Jensen C
G3 (Bethesda). 2022 Jul 28. pii: 6651068. doi: 10.1093/g3journal/jkac184.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 159821 | pSEM246 - MCS mosTI unc-119 target | Single copy or array insertion by MosTI using split unc-119 selection |
| 159822 | pSEM318 - sgRNA mosTI unc-119 single-copy | Single copy or array insertion by MosTI using split unc-119 selection |
| 159823 | pSEM319 - array co-injection fragment mosTI unc-119 | Single copy or array insertion by MosTI using split unc-119 selection |
| 159824 | pSEM320 - sgRNA mosTI unc-119 array insertion | Single copy or array insertion by MosTI using split unc-119 selection |
| 159825 | pCFJ2560 - pEXP(mosTI, unc-119)[Peft-3 | ceGFP(PATC, NLS) | tbb-2 3'UTR] | Single copy or array insertion by MosTI using split unc-119 selection |
| 159826 | pSEM270 - MCS mosTI hygroR target | Single copy or array insertion by MosTI using split hygroR selection |
| 159827 | pSEM253 - sgRNA mosTI hygroR | Single copy or array insertion by MosTI using split hygroR selection |
| 159828 | pSEM321 - array co-injection fragment mosTI hygroR | Single copy or array insertion by MosTI using split hygroR selection |
| 159829 | pSEM266 - pEXP(mosTI, hygroR)[Pmex-5 | ceGFP(PATC, NLS) | tbb-2 3'UTR] | Single copy or array insertion by MosTI using split hygroR selection |
| 159830 | pSEM271 - MCS mosTI NeoR target | Single copy or array insertion by MosTI using split NeoR selection |
| 159831 | pSEM254 - sgRNA mosTI neoR | Single copy or array insertion by MosTI using split NeoR selection |
| 159832 | pSEM322 - array co-injection fragment mosTI neoR | Single copy or array insertion by MosTI using split NeoR selection |
| 159833 | pSEM267 - pEXP(mosTI, neoR)[Pmex-5 | ceGFP(PATC, NLS) | tbb-2 3'UTR] | Single copy or array insertion by MosTI using split NeoR selection |
| 159834 | pSEM272 - MCS mosTI Pmlc-2_GFP target | Single copy or array insertion by MosTI using split Pmlc-2 GFP selection |
| 159835 | pSEM255 - sgRNA mosTI Pmlc-2_GFP | Single copy or array insertion by MosTI using split Pmlc-2 GFP selection |
| 159836 | pSEM323 - array co-injection fragment mosTI Pmlc-2_GFP | Single copy or array insertion by MosTI using split Pmlc-2 GFP selection |
| 159840 | pSEM262 - pDESTR4-R3 mosTI hygroR target | Single copy or array insertion by MosTI using split hygroR selection |
| 159842 | pSEM264 - pDESTR4-R3 mosTI Pmlc-2_GFP target | Single copy or array insertion by MosTI using split Pmlc-2 GFP selection |
| 182343 | pSEM246 - mosTI - unc-119 - MCS | Empty cloning vector for mosTI(unc-119). MCS or Golden Gate (BsaI: tgcc - MCS - gagc) |
| 182344 | pSEM376 - sgRNA - spc4 - array - insertion - unc-119 - locus | sgRNA4 for insertion of extrachromosomal arrays into the ce-unc-119 genomic locus. Use with pSEM371 fragment included in Ex array. |
| 182345 | pSEM386 - mosTI unc-119 - NNN - gfp - tbb-2 | GFP fusion cloning vector for mosTI(unc-119). Transgene inserted upstream of gfp and tbb-2 3' UTR. MCS or Golden Gate (BsaI: tgcc - MCS - aaaa) |
| 182347 | pSEM387 - mosTI unc-119 - NNN - gfp - nls - tbb-2 | GFP transcriptional reporter cloning vector for mosTI(unc-119). Transgene inserted upstream of gfp(2xNLS) and tbb-2 3' UTR. MCS or Golden Gate (BsaI: tgcc - MCS - aaaa) |
| 182348 | pSEM388 - mosTI unc-119 - NNN - mScarlet - tbb-2 | mScarlet fusion cloning vector for mosTI(unc-119). Transgene inserted upstream of mScarlet and tbb-2 3' UTR. MCS or Golden Gate (BsaI: tgcc - MCS - aaaa) |
| 182349 | pSEM389 - mosTI unc-119 - NNN - mScarlet(nls) - tbb-2 | mScarlet transcriptional reporter cloning vector for mosTI(unc-119). Transgene inserted upstream of mScarlet(2xNLS) and tbb-2 3' UTR. MCS or Golden Gate (BsaI: tgcc - MCS - aaaa) |
| 182350 | pSEM390 - mosTI unc-119 - Peft-3 - NNN - tbb-2 | Ubiquitous expression cloning vector for mosTI(unc-119). Transgene inserted between Peft-3 and tbb-2 3'UTR. MCS or Golden Gate (BsaI: aaaa - MCS - tgag) |
| 182351 | pSEM391 - mosTI unc-119 - Pmex-5 - NNN - tbb-2 | Germline expression cloning vector for mosTI(unc-119). Transgene inserted between Pmex-5 and tbb-2 3'UTR. MCS or Golden Gate (BsaI: aaaa - MCS - tgag) |
| 182352 | pSEM394 - mosTI unc-119 - PUPN - NNN - tbb-2 | Pan-neuronal expression cloning vector for mosTI(unc-119). Transgene inserted between Ultra pan neuronal (UPN) promoter and tbb-2 3'UTR. MCS or Golden Gate (BsaI: aaaa - MCS - tgag) |
| 182353 | pSEM396 - mosTI unc-119 - NNN - operon - gfp - tbb-2 | GFP co-expression cloning vector for mosTI(unc-119). Transgene inserted upstream of gpd-2 operon::gfp::tbb-2 3'UTR. MCS or Golden Gate (BsaI: tgcc - MCS - tgag) |
| 182354 | pSEM397 - mosTI unc-119 - NNN - operon - gfp - nls - tbb-2 | Nuclear GFP co-expression cloning vector for mosTI(unc-119). Transgene inserted upstream of gpd-2 operon::gfp(2xNLS)::tbb-2 3'UTR. MCS or Golden Gate (BsaI: tgcc - MCS - tgag) |
| 182355 | pSEM398 - mosTI unc-119 - NNN - operon - mScarlet - tbb-2 | mScarlet co-expression cloning vector for mosTI(unc-119). Transgene inserted upstream of gpd-2 operon::mScarlet::tbb-2 3'UTR. MCS or Golden Gate (BsaI: tgcc - MCS - tgag) |
| 182356 | pSEM399 - mosTI unc-119 - NNN - operon - mScarlet - nls - tbb-2 | Nuclear mScarlet co-expression cloning vector for mosTI(unc-119). Transgene inserted upstream of gpd-2 operon::mScarlet(2xNLS)::tbb-2 3'UTR. MCS or Golden Gate (BsaI: tgcc - MCS - tgag) |
| 182357 | pSEM414 - mosTI unc-119 Peft-3 - gfp(nls) - tbb-2 | Positive control for mosTI(unc-119). unc-119 rescue and ubiquitous nuclear GFP expression after mosTI insertion. |
| 182358 | pSEM415 - mosTI unc-119 Pmex-5 - gfp(nls) - tbb-2 | Positive control for mosTI(unc-119). unc-119 rescue and germline nuclear GFP expression after mosTI insertion. |
| 182359 | pSEM416 - mosTI unc-119 P_UPN - gfp(nls) - tbb-2 | Positive control for mosTI(unc-119). unc-119 rescue and pan-neuronal nuclear GFP expression after mosTI insertion. |
| 182640 | pSEM371 - ce-unc-119 array integration | Co-injection fragment for extra-chromosomal array integration at ce-unc-119 locus |
| 190933 | pCFJ782 - HygroR | Hygromycin resistance co-injection marker |
| 191381 | pMDJ39 - Peft-3 Cre (PATCs) | High efficiency CRE recombinase expression in C. elegans |
| 191382 | pMDJ231 - Phsp Cas9 | Heat shock inducible Cas9 expression in C. elegans. |