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Knockout of microRNAs using Cas9 nuclease and synthetic crRNA:tracrRNA
Poster Title: Knockout of microRNAs using Cas9 nuclease and synthetic crRNA:tracrRNA
Submitted on 22 Jan 2016
Author(s): Eldon T. Chou, John A. Schiel, Elena Maksimova, Emily M. Anderson, Melissa L. Kelley, and Anja van Brabant Smith | Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US
Affiliations: Dharmacon (part of GE Healthcare)
This poster was presented at Keystone 2016
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Abstract: The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated proteins) system derived from Streptococcus pyogenes uses a Cas9 nuclease protein guided by two small RNA sequences, the tracrRNA and a targeting crRNA containing a 20 nucleotide guide sequence complementary to the genomic target of interest, to create double-strand DNA breaks (Figure 1). The double-strand DNA break is repaired by either non-homologous end joining (NHEJ) or homology-directed repair (HDR) by endogenous mechanisms within the mammalian cells. NHEJ often results in small insertions or deletions (indels) that produce functional gene knockouts through nonsense mutations or introduction of a stop codon. Summary: Using Edit-R Cas9 plasmid with custom synthetic crRNAs and tracrRNA to knock out endogenous microRNAs.Report abuse »
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