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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. PubMed Article

Plasmids from Article

ID Plasmid Purpose
159821pSEM246 - MCS mosTI unc-119 targetSingle copy or array insertion by MosTI using split unc-119 selection
159822pSEM318 - sgRNA mosTI unc-119 single-copySingle copy or array insertion by MosTI using split unc-119 selection
159823pSEM319 - array co-injection fragment mosTI unc-119Single copy or array insertion by MosTI using split unc-119 selection
159824pSEM320 - sgRNA mosTI unc-119 array insertionSingle copy or array insertion by MosTI using split unc-119 selection
159825pCFJ2560 - 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
159826pSEM270 - MCS mosTI hygroR targetSingle copy or array insertion by MosTI using split hygroR selection
159827pSEM253 - sgRNA mosTI hygroRSingle copy or array insertion by MosTI using split hygroR selection
159828pSEM321 - array co-injection fragment mosTI hygroRSingle copy or array insertion by MosTI using split hygroR selection
159829pSEM266 - pEXP(mosTI, hygroR)[Pmex-5 | ceGFP(PATC, NLS) | tbb-2 3'UTR]Single copy or array insertion by MosTI using split hygroR selection
159830pSEM271 - MCS mosTI NeoR targetSingle copy or array insertion by MosTI using split NeoR selection
159831pSEM254 - sgRNA mosTI neoRSingle copy or array insertion by MosTI using split NeoR selection
159832pSEM322 - array co-injection fragment mosTI neoRSingle copy or array insertion by MosTI using split NeoR selection
159833pSEM267 - pEXP(mosTI, neoR)[Pmex-5 | ceGFP(PATC, NLS) | tbb-2 3'UTR]Single copy or array insertion by MosTI using split NeoR selection
159834pSEM272 - MCS mosTI Pmlc-2_GFP targetSingle copy or array insertion by MosTI using split Pmlc-2 GFP selection
159835pSEM255 - sgRNA mosTI Pmlc-2_GFPSingle copy or array insertion by MosTI using split Pmlc-2 GFP selection
159836pSEM323 - array co-injection fragment mosTI Pmlc-2_GFPSingle copy or array insertion by MosTI using split Pmlc-2 GFP selection
159840pSEM262 - pDESTR4-R3 mosTI hygroR targetSingle copy or array insertion by MosTI using split hygroR selection
159842pSEM264 - pDESTR4-R3 mosTI Pmlc-2_GFP targetSingle copy or array insertion by MosTI using split Pmlc-2 GFP selection
182343pSEM246 - mosTI - unc-119 - MCSEmpty cloning vector for mosTI(unc-119). MCS or Golden Gate (BsaI: tgcc - MCS - gagc)
182344pSEM376 - sgRNA - spc4 - array - insertion - unc-119 - locussgRNA4 for insertion of extrachromosomal arrays into the ce-unc-119 genomic locus. Use with pSEM371 fragment included in Ex array.
182345pSEM386 - mosTI unc-119 - NNN - gfp - tbb-2GFP 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)
182347pSEM387 - mosTI unc-119 - NNN - gfp - nls - tbb-2GFP 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)
182348pSEM388 - mosTI unc-119 - NNN - mScarlet - tbb-2mScarlet 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)
182349pSEM389 - mosTI unc-119 - NNN - mScarlet(nls) - tbb-2mScarlet 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)
182350pSEM390 - mosTI unc-119 - Peft-3 - NNN - tbb-2Ubiquitous 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)
182351pSEM391 - mosTI unc-119 - Pmex-5 - NNN - tbb-2Germline 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)
182352pSEM394 - mosTI unc-119 - PUPN - NNN - tbb-2Pan-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)
182353pSEM396 - mosTI unc-119 - NNN - operon - gfp - tbb-2GFP 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)
182354pSEM397 - mosTI unc-119 - NNN - operon - gfp - nls - tbb-2Nuclear 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)
182355pSEM398 - mosTI unc-119 - NNN - operon - mScarlet - tbb-2mScarlet 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)
182356pSEM399 - mosTI unc-119 - NNN - operon - mScarlet - nls - tbb-2Nuclear 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)
182357pSEM414 - mosTI unc-119 Peft-3 - gfp(nls) - tbb-2Positive control for mosTI(unc-119). unc-119 rescue and ubiquitous nuclear GFP expression after mosTI insertion.
182358pSEM415 - mosTI unc-119 Pmex-5 - gfp(nls) - tbb-2Positive control for mosTI(unc-119). unc-119 rescue and germline nuclear GFP expression after mosTI insertion.
182359pSEM416 - mosTI unc-119 P_UPN - gfp(nls) - tbb-2Positive control for mosTI(unc-119). unc-119 rescue and pan-neuronal nuclear GFP expression after mosTI insertion.
182640pSEM371 - ce-unc-119 array integrationCo-injection fragment for extra-chromosomal array integration at ce-unc-119 locus
190933pCFJ782 - HygroRHygromycin resistance co-injection marker
191381pMDJ39 - Peft-3 Cre (PATCs) High efficiency CRE recombinase expression in C. elegans
191382pMDJ231 - Phsp Cas9Heat shock inducible Cas9 expression in C. elegans.

Antibodies from Article