Therefore, avoid using the reagents which have been previously frozen. Freezing can deteriorate the reagents, which can produce inaccurate results. Precautions for use 1) This product should be stored as directed, without freezing. High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. ample water as first aid, and consult the doctor if required. Correction of a pathogenic gene mutation in human embryos. DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins. Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery. Genome-wide target specificities of CRISPR RNA-guided programmable deaminases. Digenome-seq: genome-wide profiling of CRISPR-Cas9 off-target effects in human cells. Functional correction of large factor VIII gene chromosomal inversions in hemophilia a patient-derived iPSCs using CRISPR-Cas9. Targeted chromosomal duplications and inversions in the human genome using zinc finger nucleases. Targeted chromosomal deletions in human cells using zinc finger nucleases. Targeted AID-mediated mutagenesis (TAM) enables efficient genomic diversification in mammalian cells. Directed evolution using dCas9-targeted somatic hypermutation in mammalian cells. Functional footprinting of regulatory DNA. CRISPR-mediated base editing enables efficient disruption of eukaryotic genes through induction of STOP codons. CRISPR-STOP: gene silencing through base-editing-induced nonsense mutations. Programmable base editing of A*T to G*C in genomic DNA without DNA cleavage. Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C:G-to-T:A base editors with higher efficiency and product purity.
Enhanced base editing by co-expression of free uracil DNA glycosylase inhibitor. Highly efficient RNA-guided base editing in mouse embryos. Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion. Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. A guide to genome engineering with programmable nucleases. The results "strongly suggest that mesophiles and thermophiles fundamentally differ in their use of adenine for this electron transfer repair mechanism," Rousseau said.Kim, H. coli, the adenine was in a different position, providing mainly structural support. He found that in thermophiles, adenine played a role in transferring the electron to the DNA. Coli that thrive at moderate temperatures, called mesophiles. He looked at photolyase in both the heat-loving ancestors of ancient bacteria, called thermophiles, and more modern bacteria like E. Rousseau studied the role of a molecule called adenine in shuttling the electron from the molecular antenna to the DNA strand. Credit: Benjamin Rousseau, courtesy of the Journal of the American Chemical Society The adenine-containing structure in the middle hands the electron to the DNA strand, splitting apart DNA bases. Photolyase proteins use a molecular antenna (green, blue and red structure on the right) to harvest light and convert it into an electron. It then hands the electron over to the DNA strand, sparking a reaction that splits the two bases apart and restores the genetic information. Photolyase uses a molecular antenna to capture light from the sun and convert it into an electron. UV light can trigger two adjacent bases to react and latch onto one other, rendering these genetic instructions unreadable. 28 issue of the Journal of the American Chemical Society.ĭNA is built of chains of bases-A, C, G and T-whose order encodes our genetic information. In a new study, Rousseau and coworkers, working with Professor David Beratan and Assistant Research Professor Agostino Migliore, used computer simulations to study photolyase in thermophiles, the great great great great grandchildren of Earth's original bacterial pioneers. Though these proteins have been around for billions of years, scientists are still not quite sure exactly how they work. "They are one of the most ancient repair proteins." "Anything under the sun-in both meanings of the phrase-has to have ways to repair itself, and photolyase proteins are one of them," Rousseau said.
This molecule, called photolyase, fixes DNA damaged by ultraviolet (UV) radiation-the same wavelengths of sunlight that give us sunburn and put us at greater risk of skin cancer. Benjamin Rousseau, a research technician in David Beratan's lab at Duke, studies one of the molecular machines that helped these bacteria survive their harsh environment.