Experimental design considerations for efficient and specific gene knockin using a CRISPR-Cas9 for HDR with synthetic crRNA and tracrRNA
Poster Aug 19, 2016
Hidevaldo B. Machado, John A. Schiel, Maren Mayer-Gross, Eldon T. Chou, Melissa L. Kelley, Anja van Brabant Smith. Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Lafayette, CO 80026, USA
The bacterial CRISPR-Cas9 system has revolutionized the genome engineering world with its efficiency and ease of use. The most common use of the CRISPR-Cas9 technology has been to engender gene knockouts that are generated as a result of imperfect repair of a targeted double-strand DNA break by the non-homologous end joining (NHEJ) pathway.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE
Genome-wide association studies (GWAS) have identified more than 100 genetic loci associated with type 2 diabetes. The majority of these are located in the intergenic or intragenic regions suggesting that the implicated variants may alter chromatin conformation. This, in turn, is likely to influence the expression of nearby or more remotely located genes to alter beta cell function. At present, however, detailed molecular and functional analyses are still lacking for most of these variants. We recently analysed one of these loci and mapped five causal variants in an islet-specific enhancer cluster within the STARD10 gene locus. Here, we aimed to understand how these causal variants influence b-cell function by alteration of the chromatin structure of enhancer clusterREAD MORE