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7 záznamů v Medvik Filtry

Článek

Response to correspondence on "Reproducibility of CRISPR-Cas9 methods for generation of conditional mouse alleles: a multi-center evaluation"

Gurumurthy, Channabasavaiah B
Autor Gurumurthy, Channabasavaiah B Mouse Genome Engineering Core Facility, Vice Chancellor for Research Office, University of Nebraska Medical Center, Omaha, NE, USA. cgurumurthy@unmc.edu Developmental Neuroscience, Munroe Meyer Institute for Genetics and Rehabilitation, University of Nebraska Medical Center, Omaha, NE, USA. cgurumurthy@unmc.edu
O'Brien, Aidan R
Autor O'Brien, Aidan R Transformational Bioinformatics, Health and Biosecurity Business Unit, CSIRO, Sydney, Australia Department of Immunology and Infectious Disease, the John Curtin School of Medical Research, The Australian National University, Canberra, Australia
Quadros, Rolen M
Autor Quadros, Rolen M Mouse Genome Engineering Core Facility, Vice Chancellor for Research Office, University of Nebraska Medical Center, Omaha, NE, USA
Adams, John
Autor Adams, John Texas A&M Institute for Genomic Medicine (TIGM), Texas A&M University, College Station, TX, 77843, USA
Alcaide, Pilar
Autor Alcaide, Pilar Department of Immunology, Tufts University School of Medicine, Boston, USA
Ayabe, Shinya
Autor Ayabe, Shinya RIKEN BioResource Research Center, Tsukuba, Ibaraki, 305-0074, Japan
Ballard, Johnathan
Autor Ballard, Johnathan Texas A&M Institute for Genomic Medicine (TIGM), Texas A&M University, College Station, TX, 77843, USA
Batra, Surinder K
Autor Batra, Surinder K Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE, USA
Beauchamp, Marie-Claude
Autor Beauchamp, Marie-Claude Departments of Anatomy and Cell Biology, Human Genetics and Pediatrics, Research Institute McGill University Health Center (RI-MUHC), Montreal, Canada
Becker, Kathleen A
Autor Becker, Kathleen A Maine Medical Center Research Institute (MMCRI), Scarborough, ME, USA

Genome biology. 2021 ; 22 (1) : 99. [pub] 20210407

Genome Biol
ISSN 1474-760X
Medvik
Zdroj

Článek

Reproducibility of CRISPR-Cas9 methods for generation of conditional mouse alleles: a multi-center evaluation

Genome biology. 2019 ; 20 (1) : 171. [pub] 20190826

Genome Biol
ISSN 1474-760X
Medvik
Zdroj

BACKGROUND: CRISPR-Cas9 gene-editing technology has facilitated the generation of knockout mice, providing an alternative to cumbersome and time-consuming traditional embryonic stem cell-based methods. An earlier study reported up to 16% efficiency in generating conditional knockout (cKO or floxed) alleles by microinjection of 2 single guide RNAs (sgRNA) and 2 single-stranded oligonucleotides as donors (referred herein as "two-donor floxing" method). RESULTS: We re-evaluate the two-donor method from a consortium of 20 laboratories across the world. The dataset constitutes 56 genetic loci, 17,887 zygotes, and 1718 live-born mice, of which only 15 (0.87%) mice contain cKO alleles. We subject the dataset to statistical analyses and a machine learning algorithm, which reveals that none of the factors analyzed was predictive for the success of this method. We test some of the newer methods that use one-donor DNA on 18 loci for which the two-donor approach failed to produce cKO alleles. We find that the one-donor methods are 10- to 20-fold more efficient than the two-donor approach. CONCLUSION: We propose that the two-donor method lacks efficiency because it relies on two simultaneous recombination events in cis, an outcome that is dwarfed by pervasive accompanying undesired editing events. The methods that use one-donor DNA are fairly efficient as they rely on only one recombination event, and the probability of correct insertion of the donor cassette without unanticipated mutational events is much higher. Therefore, one-donor methods offer higher efficiencies for the routine generation of cKO animal models.

MeSH
alely * MeSH
blastocysta metabolismus MeSH
CRISPR-Cas systémy genetika MeSH
faktorová analýza statistická MeSH
mikroinjekce MeSH
myši knockoutované MeSH
protein 2 vázající methyl-CpG genetika metabolismus MeSH
protein Cas9 metabolismus MeSH
regresní analýza MeSH
reprodukovatelnost výsledků MeSH
zvířata MeSH
Check Tag
mužské pohlaví MeSH
ženské pohlaví MeSH
zvířata MeSH
Publikační typ
časopisecké články MeSH
multicentrická studie MeSH
práce podpořená grantem MeSH
Research Support, N.I.H., Extramural MeSH

Článek

Overall Survival Analysis From a Randomized Phase II Study of Axitinib With or Without Dose Titration in First-Line Metastatic Renal Cell Carcinoma

Clinical genitourinary cancer. 2016 ; 14 (6) : 499-503. [pub] 20160422

Clin Genitourin Cancer
ISSN 1938-0682
Medvik
Zdroj

BACKGROUND: In a randomized phase II trial in metastatic renal cell carcinoma (mRCC), objective response rate was significantly higher with axitinib versus placebo titration (54% vs. 34%; 1-sided P = .019). PATIENTS AND METHODS: Treatment-naive patients with mRCC (n = 213) received axitinib 5 mg twice per day (b.i.d.) for 4 weeks. Patients meeting dose titration criteria were randomized to receive axitinib 5 mg b.i.d. with axitinib or placebo titration (n = 56 each); 91 patients ineligible for randomization continued axitinib 5 mg b.i.d.; 10 discontinued before randomization. RESULTS: Median overall survival (95% confidence interval [CI]) was 42.7 months (24.7-not estimable) with axitinib titration versus 30.4 months (23.7-45.0) with placebo titration (stratified hazard ratio, 0.785; 95% CI, 0.485-1.272; 1-sided P = .162), and 41.6 months (95% CI, 33.0-not estimable) in nonrandomized patients. Safety data were consistent with previous reports. CONCLUSION: Median overall survival was numerically longer in patients with first-line mRCC who received axitinib versus placebo titration. No new safety signal was observed after long-term axitinib treatment in first-line mRCC.