Bolisetty MT, Beemon KL. (2012) Splicing of internal large exons is defined by novel cis-acting sequence elements. Nucleic Acids Res, Epub Ahead of Print(0):1–11.
In this paper by Bolisetty and Beemon, the researchers investigated splicing regulation of large exons, >1000 nt. They identified 18 sites in the 1039 nt exon 7 of the gene JARID2 that contained a consensus sequence that is “C-rich with a central invariant CA dinucleotide” that they hypothesized was important for proper splicing. In order to confirm their hypothesis, the authors needed to mutate all 18 of the identified sites. Typically, creating a large construct with this many mutations would involve multiple mutagenesis reactions, or assembling a large number of synthetic oligonucleotides. However, Bolisetty and Beemon required only 3 synthetic, high fidelity, double-stranded gBlocks® Gene Fragments from IDT to cover the entire exon 7 sequence, with all 18 mutated sites. The ability to order entire sequences up to 3000 bp that are ready for easy assembly demonstrates the exceptional versatility and timesaving ability of gBlocks Gene Fragments.
gBlocks® Gene Fragments are double-stranded, sequence-verified, DNA genomic blocks, 125–3000 bp in length, that can be shipped in 2–5 working days for affordable and easy gene construction or modification. gBlocks Gene Fragments have been used in a wide range of applications including CRISPR-mediated genome editing, antibody research, codon optimization, mutagenesis, and aptamer expression. They can also be used for generating qPCR standards. With gBlocks Gene Fragments, IDT provides a tool that accelerates your research and makes synthetic biology easier and more accessible than ever, for any lab.
Learn more about gBlocks Gene Fragments at www.idtdna.com/gblocks.
Read how gBlocks Gene Fragments have been applied in other research projects:
Towards next generation biosensors—See how fluorescent protein-based Ca2+ indicators based on naturally occurring substrates are assembled using gBlocks Gene Fragments. These sensors are being developed to monitor in vivo neural activity.
Site-directed mutagenesis—improvements to established methods—Site-directed mutagenesis techniques have relied primarily on PCR and standard cloning methods. Read about some of the common cloning methods used for mutagenesis and how double-stranded DNA fragments (gBlocks Gene Fragments) can save you both time and money.
Easily-Designed Standard Curves for qPCR—Use this easy way to combine control templates/multiple targets onto a single construct, and the advantages that it can provide for PCR experiments.
Review other DECODED Online newsletter articles on mutagenesis or cloning and synthetic biology applications.
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