The CRISPR-Cas type V-I is a family of Cas12i-containing programmable nuclease systems guided by a short crRNA without requirement for a tracrRNA. Here we present an engineered Type V-I CRISPR system (Cas12i), ABR-001, which utilizes a tracr-less guide RNA. The compact Cas12i effector is capable of self-processing pre-crRNA and cleaving dsDNA targets, which facilitates versatile delivery options and multiplexing, respectively.
View Article and Find Full Text PDFCell-free protein synthesis is a powerful tool to produce recombinant proteins, and as an open system, can often integrate all or part of downstream assays. Here we describe in vitro synthesis of the Streptococcus pyogenes type II-A CRISPR-Cas9 ribonucleoproteins (SpCas9 RNPs), consisting of the effector protein and the single guide RNAs (sgRNAs). In spite of its large molecular weight (160kDa), the SpCas9 effector is expressed relatively well from linear DNA templates under T7 promoter in commercial reconstituted cell-free protein synthesis systems.
View Article and Find Full Text PDFDNA polymerases have revolutionized the biotechnology field due to their ability to precisely replicate stored genetic information. Screening variants of these enzymes for specific properties gives the opportunity to identify polymerases with different features. We have previously developed a single-molecule DNA sequencing platform by coupling a DNA polymerase to an α-hemolysin pore on a nanopore array.
View Article and Find Full Text PDFBackground And Study Aims: Barrett's Esophagus (BE) is a well-recognized pre-malignant condition. Previous data indicate histologically confirmed BE frequency varies by ethnicity in the United States. However, clinical factor assessment to explain this has only occurred in a veteran population to date.
View Article and Find Full Text PDFAim: To evaluate if differences exist between self-expanding esophageal metal stents (SEMS) and self-expanding esophageal plastic stents (SEPS) when used for benign or malignant esophageal disorders with regard to safety, efficacy, clinical outcomes, placement ease and cost.
Methods: A retrospective analysis was performed to evaluate outcome in patients having SEPS/SEMS placed for malignant or benign esophageal conditions from January 2005 to April 2012. Inclusion criteria was completed SEMS/SEPS placement.