rRMCE landing sites

Rapid RMCE landing sites have a general structure consisting of two transcription units flanked by a 5′ loxP site and a 3′ FRT3 site. The first consists of a promoter driving a Orange Fluorescent Protein (cyOFP) with an FRT site positioned at the junction between the promoter and the FP (in most cases). This permits visualization of the landing site and easy detection of novel insertions. The second transcription unit consists of FLP D5 driven by the mex-5 promoter and drives the recombination reaction. This portion of the landing site is excised during the recombination reaction.

The following landing sites are all described in Nonet (2023).

Chromosomepharynx labelled (myo-2p FRT
nls-cyOFP)		TRNs labelled
(mec-4p FRT
nls-cyOFP)		CCs labelled
(cup-4p FRT cyOFP)		unlabelled*
(FRT myo-2p
nls-cyOFP)
I
at jsTi1453		jsSi1727jsSi1971jsSi1962
II
near ttTi5605		jsSi1726jsSi1944jsSi1900jsSi1901
IV
near cxTi10882		jsSi1986jsSi1837jsSi1988jsSi1963
V
near oxTi365		jsSi1985jsSi1987jsSi1978
Table 1. Landing sites for rRMCE. Landing sites in this table are organized by the visual marker that will be associated with the final transgene (top column). All landing site are marked with cyOFP expressed under in the cell type listed (or myo-2p in the case of unmarked transgenes). Click on the allele for the sequence of the insertion which were determined by nano-pore sequencing of PCR products that span the insertion site. TRNs= touch receptor neurons, CC= coelomocytes. *= The landing site is marked with myo-2p nls-cyOFP, but the myo-2p and cyOFP are excised during integration. All strains are available from the CGC.

If none of these sites suites your needed, one can build novel landing sties using RMCE or CRISPR (see Nonet, 2023 for details).

Nonet, M. (2023). Rapid generation of C. elegans single copy transgenes combining RMCE and drug selection.  Genetics iyad072. PMID:37079426 [Abstract] [PDF]

Last edited 6-28-2023