This study reports a DNA-free genome editing method in potato via delivery of Alt-R CRISPR-Cas9 ribonucleoprotein (RNP) to potato protoplasts. The authors demonstrate that RNP delivery of CRISPR-Cas9 components comprising synthetic RNA yields higher frequencies of transgene-free mutated lines compared to using in vitro transcribed RNA or plasmid DNA delivery. Therefore, they propose using RNP with synthetic RNAs for CRISPR in potato plants to simplify analysis and selection of commercial crop lines.
In an example of the evolution of DNA-based nanomachinery, researchers devise 2 anti-parallel DNA origami filaments that undergo step-wise, reversible sliding in opposite directions. Movement is powered by DNA fuels mediated by gold nanocrystals.
This study describes a robust and simple-to-perform method called improved-Genome editing via Oviductal Nucleic Acids Delivery (i-GONAD), which delivers CRISPR RNP to E0.7 embryos via in situ electroporation.
This study reports dysregulated blood glucose homeostasis in cave-adapted populations of the Mexican tetra, Astyanax mexicanus, and suggests a beneficial effect of diminished insulin signaling in a nutrient-limited environment. The researchers used the 2-part Alt-R guide RNAs from IDT to introduce cavefish-specific point mutations into zebrafish and study the gain-of-function phenotype.
PrimeTime Assays using FAM-labeled, ZEN Double-Quenched Probes were used to quantify total mitochondrial DNA in mice.
ZEN Double-Quenched Probes were used to increase the sensitivity of real-time RT-PCR assays designed for analysis of RotaTeq virus. The assays were designed to analyze 5 genotypes (G1, G2, G3, G4, and G6) in the VP7 gene.
This study from Genentech describes an optimized method for delivering CRISPR-Cas9 RNP into primary mouse and human T cells. The use of Cas9 RNP transfection overcomes the limitations of all-in-one lentiviral approaches and does not require stimulation of TCR (T cell receptor), thus allowing functional studies of genes involved in T cell activation and differentiation. The researchers achieved highly efficient gene knock-out via nucleofection of a Cas9 RNP formed by Alt-R crRNA and tracrRNA, which largely mitigates the need for selection and clonal isolation.
Researchers from Genentech use IDT Alt-R CRISPR RNAs to efficiently knock out endogenous genes RIPK1 and HOIP in Bone Marrow-Derived Macrophages (BMDM).
The researchers address the incorporation and accessiblitiy of DNA strands in DNA origami nanostructures with molecular resolution using DNA-PAINT super resolution microscopy. This method can provide feedback to refine design and assembly of these structures.
The authors demonstrate that sample cross talk due to combinatorial indexes can be corrected by using unique adapters containing unique molecular identifiers (UMIs). Their results suggest that such adapters are critical to reducing false positive rates in sensitive applications, such as somatic variant discovery. The adapter design presented in this article uses dual matched indexes. xGen Dual Index UMI Adapters—Tech Access incorporate an updated design, using dual unique indexes based on Illumina’s best practice guidelines.
Here, researchers deliver Cas9 ribonucleoprotein (RNP) to various cell types, including human primary CD4+ T cells, via a novel microfluidic cell deformation-based method.
The authors use DNA origami with GFP antibodies to provide a system for calibrating fluorophore and antibody labeling efficiency. The method should allow researchers to quantify protein copy number within cells using super-resolution microscopy.
This study shows that in vitro-assembled, dual Alt-R Cas9 RNPs coupled with microhomology repair templates enable efficient gene manipulation in different genetic backgrounds of A. fumigatus.
This publication reports a screening effort to uncover novel regulators of vertebrate spindle orientation. The authors describe a method for efficient gene knockout in chick embryos which employs in ovo electroporation of Alt-R CRISPR-Cas9 components (crRNA and tracrRNA).
xGen Lockdown Probes and Blocking Oligos were used to generate a custom gene panel that targeted 382 cancer-relevant genes in multiple cancer types. The panel provided target enrichment across 605 circulating tumor (ctDNA) samples.
Exome sequencing with the xGen Exome Research Panel established a molecular genetic diagnosis of TARP syndrome for a neonatal patient in whom traditional testing methods were uninformative. Sequencing of the proband, mother, and father showed a previously unreported, maternally-inherited alteration in an RNA-binding motif protein.This allowed efficient diagnosis and future reproductive options for the parents.
26 paired samples (FFPE-derived tumor samples + frozen, PMBC-derived normal samples) were sequenced by exome hybrid capture using the xGen Exome Research Panel. RNA sequencing was also performed using the panel. The study demonstrates that improvements to the efficacy of immune-mobilizing therapies require the genetic profiling of both the tumor and immune system.