Publication of Methodology Useful for Research in DM

Methods in Molecular Biology is a long-running series disseminating research protocols and methodologies. A strength is the ability to precisely reproduce methodologies based upon step-by-step protocols.

In the latest release of Methods in Molecular Biology, six protocols from the myotonic dystrophy field are included. These include:

• FISH techniques to detect expanded CUG repeat RNA or DMPK transcripts in DM1 plus a combined FISH/immunofluorescence protocol to visualize the colocalization of expanded repeat RNA and MBNL1 (Klein et al., 2020);
• Methodology for in vitro synthesis and structure analysis of expanded CUG repeat RNA (van Cruchten and Wansink, 2020);
• Technology for a tractable cell culture system to study instability of expanded CUG repeats; (Kononenko et al., 2020);
• Use of FLP recombinase to construct HeLa clonal cell lines containing CNG repeats of various lengths at a single ectopic recombination acceptor site (Gadgil et al., 2020);
• Design of PCR-based assays, using the model system Saccharomyces cerevisiae, to determine the behavior of expanded CTG tracts in stable and DM1 threshold ranges (Williams et al., 2020); and
• Protocols for expanded repeat-primed PCR and restriction enzyme-digested PCR to detect/identify interrupted expanded repeat alleles in DM1 (Tomé and Gourdon, 2020).

MDF is including this item in the Research News to highlight the availability of these now-published research protocols.

References:

FISH Protocol for Myotonic Dystrophy Type 1 Cells.
Klein AF, Arandel L, Marie J, Furling D.
Methods Mol Biol. 2020;2056:203-215. doi: 10.1007/978-1-4939-9784-8_13.

In Vitro Synthesis and RNA Structure Probing of CUG Triplet Repeat RNA.
van Cruchten RTP, Wansink DG.
Methods Mol Biol. 2020;2056:187-202. doi: 10.1007/978-1-4939-9784-8_12.

Experimental System to Study Instability of (CGG)_n Repeats in Cultured Mammalian Cells.
Kononenko AV, Ebersole T, Mirkin SM.
Methods Mol Biol. 2020;2056:137-150. doi: 10.1007/978-1-4939-9784-8_9.

Analysis of Trinucleotide Repeat Stability by Integration at a Chromosomal Ectopic Site.
Gadgil RY, Rider SD Jr, Lewis T, Barthelemy J, Leffak M.
Methods Mol Biol. 2020;2056:121-136. doi: 10.1007/978-1-4939-9784-8_8.

Tracking Expansions of Stable and Threshold Length Trinucleotide Repeat Tracts In Vivo and In Vitro Using Saccharomyces cerevisiae.
Williams GM, Petrides AK, Balakrishnan L, Surtees JA.
Methods Mol Biol. 2020;2056:25-68. doi: 10.1007/978-1-4939-9784-8_3.

Fast Assays to Detect Interruptions in CTG.CAG Repeat Expansions.
Tomé S, Gourdon G.
Methods Mol Biol. 2020;2056:11-23. doi: 10.1007/978-1-4939-9784-8_2.