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1.
Mol Cell ; 80(2): 246-262.e4, 2020 10 15.
Article in English | MEDLINE | ID: mdl-32949493

ABSTRACT

CRISPR-Cas9-based gene drive systems possess the inherent capacity to spread progressively throughout target populations. Here we describe two self-copying (or active) guide RNA-only genetic elements, called e-CHACRs and ERACRs. These elements use Cas9 produced in trans by a gene drive either to inactivate the cas9 transgene (e-CHACRs) or to delete and replace the gene drive (ERACRs). e-CHACRs can be inserted at various genomic locations and carry two or more gRNAs, the first copying the e-CHACR and the second mutating and inactivating the cas9 transgene. Alternatively, ERACRs are inserted at the same genomic location as a gene drive, carrying two gRNAs that cut on either side of the gene drive to excise it. e-CHACRs efficiently inactivate Cas9 and can drive to completion in cage experiments. Similarly, ERACRs, particularly those carrying a recoded cDNA-restoring endogenous gene activity, can drive reliably to fully replace a gene drive. We compare the strengths of these two systems.


Subject(s)
Gene Deletion , Gene Drive Technology , Animals , CRISPR-Associated Protein 9/metabolism , Chromosomes/genetics , Drosophila melanogaster/genetics , Female , Green Fluorescent Proteins/metabolism , Inheritance Patterns/genetics , Mutagenesis/genetics , RNA, Guide, Kinetoplastida/genetics , Transgenes
2.
Pediatrics ; 122 Suppl 3: S161-70, 2008 Nov.
Article in English | MEDLINE | ID: mdl-18978010

ABSTRACT

Venipuncture and intravenous cannulation are among the most common and widespread medical procedures performed on children today. Therefore, effective treatment of venous access pain can benefit from an integrated systems approach that enlists multiple players in the health care system. By using case studies that analyze this issue from the perspective of the nurse, the physician, the pharmacist, and the child life specialist, this article illustrates how multidisciplinary programs designed to manage needle pain have been developed successfully in several institutions. Common themes that arise from these case studies include the importance of a multidisciplinary evidence-based approach to advocate change; a system-wide protocol for the administration of local anesthetics; convenient access to topical local anesthetics; department and hospital-wide support for educational efforts, including training in nonpharmacologic techniques used by child life specialists; and ongoing quantification of the overall success of any program. Implementation of these strategies can result in significant improvements in the pediatric venous access experience.


Subject(s)
Catheterization, Peripheral/adverse effects , Hospitals, Pediatric , Learning , Pain Clinics , Pain/prevention & control , Catheterization, Peripheral/trends , Disease Management , Hospitals, Pediatric/trends , Humans , Pain/etiology , Pain Clinics/trends , Pain Measurement/methods
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