|Experimental design considerations for efficient and specific gene knockin using a CRISPR-Cas9 for HDR with synthetic crRNA and tracrRNA|
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
Precise genome engineering with CRISPR-Cas9 and single-stranded DNA oligo or double-stranded DNA plasmid donors via homology-directed repair (HDR).
|CRISPR-Cas9 genome editing utilizing chemically synthesized RNA|
Kaizhang He, Eldon Chou, Amanda Haas, Žaklina Strezoska, Melissa L. Kelley, and Anja van Brabant Smith Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Lafayette, CO 80026, USA
CRISPR-Cas9 gene editing using synthetic crRNA:tracrRNA or sgRNA is highly efficient and easy to use. Synthetic crRNA:tracrRNA is uniquely suited to in vitro and in vivo applications, in particular, DNA-free approach with Cas9 mRNA. Chemical synthesis of guide RNAs allows accurate and rapid production of arrayed crRNA libraries for high-confidence, loss-of-function screens.
|Advanced Microfluidic Mixing Device for the Study of Macromolecule Dynamics|
Shubha Jain, F. Azam, Dr. H. N. Unni
We have developed and characterized a micro-fluidic mixer to study the macro-molecule dynamics such as kinetics of protein folding, DNA sequencing, single molecule study and detection etc. on a micro-second timescale. Numerical simulation has been performed to analyse the study of mixing performance of micro-fluidics channel.
|DNA-free CRISPR-Cas9 genome engineering in zebrafish|
Amanda Haas, Alex J. Blasky*, Rytis Prekeris*, John A. Schiel, Melissa L. Kelley, and Anja van Brabant Smith | Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite 100, Lafayette, CO 80026, USA *University of Colorado - Anschutz Medical Campus, Department of Cell & Developmental Biology, Denver, CO, USA
Poster describing the advantages of a DNA-free gene editing system and the application of this system in zebrafish.
|600 base reads on the Ion S5™ Next-Generation Sequencing System enables accurate HLA typing of 96 samples on one 530™ chip|
Peter B. Vander Horn, Cisilya Duncan, Jamsheed Ghadiri, Amneet Gulati, Diana Jeon, April Jung, Mindy Landes, Tommie Lincecum, Geoffrey Lowman, Vadim Mozhayskiy, Linus Ong, Xinzhan Peng, Maryam Shenasa, Prasanna Thwar
We have demonstrated that by combining improvements in templating and sequencing biochemistry we are able to sequence templates longer than 600 bases with high accuracy on an Ion S5 530 chip.
These improvements open the S5 use space to include haplotyping applications that require longer reads. As a demonstration of that, we accurately typed 96 HLA samples on one 530 chip.
|Addressing False Positive Variants Arising from Pseudogenes|
Risha Govind1,2, Sam Wilkinson1,3, Nicola Whiffin1,2, Shibu John1,2, Rachel J. Buchan1,2, Elizabeth Edwards1,2, Deborah J. Morris-Rosendahl1,3, James S. Ware1,2, P.J. Barton1,2, Stuart A. Cook1,2
Clinical genetic testing has been transformed in recent years by the introduction of Next-Generation Sequencing (NGS).
|Design considerations for highly specific and efficient synthetic crRNA molecules|
Anja van Brabant Smith, Emily M. Anderson, Shawn McClelland, Elena Maksimova, Tyler Reed, Steve Lenger, Žaklina Strezoska, Hidevaldo Machado Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US
An overview of our rational design algorithm for picking highly functional crRNA sequences in combination with comprehensive specificity analysis.
|Picking the best CRISPR-Cas9 targets for functional gene knockout: a machine learning algorithm based on both specificity and functionality|
Shawn McClelland, Emily M. Anderson, Žaklina Strezoska, Elena Maksimova, Annaleen Vermeulen, Steve Lenger, Tyler Reed, and Anja van Brabant Smith Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US
The CRISPR-Cas9 system has the potential to significantly advance basic and applied research.
|Scaffold design, function and over-expression of lentiviral-based microRNAs|
Angela Schoolmeesters, Melissa L. Kelley, Annaleen Vermeulen, Anja Smith, *Mayya Shveygert, *Xin Zhou, *Robert Blelloch Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, USA
Here we describe the strategy for scaffold design, the importance of an optimal promoter, and demonstrate gene target down-regulation from the over-expression of lentiviral microRNA mimics.
|Homology-directed repair with Dharmacon™ Edit-R™ CRISPR-Cas9 and single-stranded DNA oligos|
John A. Schiel, Eldon T. Chou, Maren Mayer, Emily M. Anderson , and Anja van Brabant Smith | Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US
Here we demonstrate how to perform lipid based transfections for homology directed repair using DharmaFECT Duo, CRISPR-Cas9 reagents and, synthetic DNA donor oligos.
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