Journal: Nature Communications

Article Title: Cre-Controlled CRISPR mutagenesis provides fast and easy conditional gene inactivation in zebrafish

doi: 10.1038/s41467-021-21427-6

Figure Lengend Snippet: a Scheme of the 3C rationale. A Cre effector construct controls the expression of a floxed Stop cassette upstream of the sequence encoding a fusion protein of Cas9 and GFP. In addition, a U6a promoter drives the constitutive expression of a gRNA targeting a gene of interest (GOI). Following exogenous or transgenic Cre supply, site-specific recombination results in the expression of Cas9-GFP. Combined with the gRNA a functional CRISPR complex is formed and mutates the target site within the gene of interest. b Scheme of the 3C gene inactivation construct targeting tyrosinase ( tyr ). The temperature-inducible hsp70l promotor drives expression of a floxed DsRed cassette. c Identification of transgenic animals expressing DsRed at 50 hpf after a heat treatment at 24 hpf. Example shown is a representative of a total of >100 heat-treated clutches from four independent 3C tyr transgenic insertions. Scale bar: 1000 µm.

Article Snippet: The fragment was subsequently ligated into pTol hsp70l:loxP-DsRed-GFP digested with SmaI and NheI replacing GFP and giving rise to pTol hsp70l:loxP-DsRed-loxP-Cas9-GFP (addgene #158962).

Techniques: Construct, Expressing, Sequencing, Transgenic Assay, Functional Assay, CRISPR

Journal: Genesis (New York, N.y. : 2000)

Article Title: GNrep mouse: A reporter mouse for front–rear cell polarity

doi: 10.1002/dvg.23299

Figure Lengend Snippet: Plasmids used for the generation of the GNrep polarity construct

Article Snippet: 12 , pJFRC172‐10XUAS‐loxP‐dSTOP‐loxP‐myr::GFP , https://www.addgene.org/32145/.

Techniques: Generated

Journal: Genetics

Article Title: Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes

doi: 10.1534/genetics.115.180679

Figure Lengend Snippet: Dual-marker cassettes facilitate tagging of genes with fluorescent protein transgenes or other epitopes. (A) Schematic of a dual-marker cassette his-72 :: GFP tagging vector. The selection cassette is housed within a constitutively spliced synthetic intron (dotted lines denote 5′ and 3′splice sites), such that after Cre -mediated excision of the cassette, the remaining loxP site will be spliced out of the mature GFP mRNA open reading frame, creating near seamless editing. (B) Fluorescent micrographs of endogenously tagged HIS-72::GFP (top) and HIS-72::mCherry (bottom). Scale bars, 100 µm.

Article Snippet: The loxP sites were designed to reside in an empty vector (loxP_genscript) or in a synthetic strongly spliced intron sequence within fluorescent protein (GFP_loxP_genscript, mCherry_loxP_genscript, GFP::3xHA_loxP_genscript) and/or 3xHA (3xHA_loxP_genscript) coding sequences. (See Table S2 for relevant information on all vectors in our toolkit and their features.)

Techniques: Marker, Plasmid Preparation, Selection

Journal: eLife

Article Title: The enteric pathogen Cryptosporidium parvum exports proteins into the cytosol of the infected host cell

doi: 10.7554/eLife.70451

Figure Lengend Snippet:

Article Snippet: Pre-made lentivirus was used to transform HCT-8 cells (RRID: CVCL_2478 ) with a loxP GFP/RGP color switch cassette (GenTarget Inc, San Diego, CA).

Techniques: Modification, Transgenic Assay, Expressing, Transfection, Construct, Isolation, Produced, In Vivo, Recombinant, Plasmid Preparation, Control, Sequencing, Luciferase, cDNA Synthesis, Software, Staining, Flow Cytometry