Search results for "crisp"
showing 10 items of 104 documents
Generation of C5aR1/C5aR2 Double Knockout Mice by CRISPR/Cas9 for Revealing Novel Aspects of Lung Injury
2019
Immunogold Labeling to Detect Streptococcus pyogenes Cas9 in Cell Culture and Tissues by Electron Microscopy
2019
Final publication is available from Mary Ann Liebert, Inc., publishers https://doi.org/10.1089/crispr.2019.0032. The CRISPR-Cas9 system is a powerful and yet precise DNA-editing tool in rapid development. By combining immunogold labeling and electron microscopy with the novel CRISPR-Cas9 system, we propose a new method to gain insight into the biology of this tool. In this study, we analyzed different Cas9-induced systems such as HEK293T cell line, murine oligodendrocyte progenitor cells, brain and liver to detect Cas9 expression by immunoelectron microscopy. Our results show that while Cas9 expression could be found in the nuclei and nucleopores of transfected HEK293T cells, in transfected…
Dimēriskas dCas9-FokI sistēmas izveidošana efektīvai zīdītāju šūnu genoma rekombinēšanai
2015
CRISPR/Cas9 sistēma nodrošina robustu un plaši pielietojamu genoma rekombinēšanas tehnoloģiju, kas paver iespēju zinātniekiem precīzi manipulēt ar specifickiem genoma elementiem, ko izmantot gēnu funkciju pētījumos bioloģijā un slimību attīstībā. Darba mērķis bija izveidot un optimizēt dCas9-FokI sistēmu, kas nodrošinātu stabilu un specifisku genoma rekombinēšanu cilvēka šūnās. Mēs piedāvājam alternatīvu tehnoloģiju, kas ir balstīta uz RNS vadītu dCas9-FokI sistēmu un piedāvā ievērojamu genoma rekombinēšanas potenciālu ar samazinātu nespecifiskās šķelšanas (off-targeting) efektu. Iegūtie dati liecina, ka šī izveidotā sistēma var tikt efektīvi izmantota genoma rekombinēšanai zīdītāju šūnās. …
Lactobacillus crispatus M247 oral administration: Is it really an effective strategy in the management of papillomavirus-infected women?
2022
Abstract Background Recent studies have shown the importance of the microbiota in women's health. Indeed, the persistence of Human Papilloma Virus (HPV)-related lesions in patients with dysbiosis can be the antechamber to cervical cancer. The aim of this study was to evaluate whether long term administration of oral Lactobacillus crispatus can restore eubiosis in women with HPV infections and hence achieve viral clearance. Methods In total, 160 women affected by HPV infections were enrolled at the Department of Gynecological Obstetrics of “San Paolo” Hospital, Italy between February 2021 and February 2022. The women were randomly assigned to two groups, one in treatment with oral Lactobacil…
XPO1E571K Mutation Modifies Exportin 1 Localisation and Interactome in B-cell Lymphoma
2020
The XPO1 gene encodes exportin 1 (XPO1) that controls the nuclear export of cargo proteins and RNAs. Almost 25% of primary mediastinal B-cell lymphoma (PMBL) and classical Hodgkin lymphoma (cHL) cases harboured a recurrent XPO1 point mutation (NM_003400, chr2:g61718472C>
ALS-linked FUS mutations confer loss and gain of function in the nucleus by promoting excessive formation of dysfunctional paraspeckles
2019
Mutations in the FUS gene cause amyotrophic lateral sclerosis (ALS-FUS). Mutant FUS is known to confer cytoplasmic gain of function but its effects in the nucleus are less understood. FUS is an essential component of paraspeckles, subnuclear bodies assembled on a lncRNA NEAT1. Paraspeckles may play a protective role specifically in degenerating spinal motor neurons. However it is still unknown how endogenous levels of mutant FUS would affect NEAT1/paraspeckles. Using novel cell lines with the FUS gene modified by CRISPR/Cas9 and human patient fibroblasts, we found that endogenous levels of mutant FUS cause accumulation of NEAT1 isoforms and paraspeckles. However, despite only mild cytoplasm…
CRISPR sequences are sometimes erroneously translated and can contaminate public databases with spurious proteins containing spaced repeats
2020
© The Author(s) 2020.
Pasquale Culotta, Crispino Valenziano e la tradizione della Composizione Cultuale nella Scuola di Architettura
2022
Lo scritto vuole compiere una verifica su alcune linee di ricerca del progetto per l’architettura cultuale nel quadro della cultura architettonica contemporanea in Sicilia a partire dall’esperienza di Pasquale Culotta e Crispino Valenziano. Dal loro sodalizio, nell’intreccio fecondo tra la dimensione speculativa negli scritti e nelle riflessioni teoriche e la sperimentazione concreta nell’architettura progettata e costruita, emerge la sistematica trattazione di una qualità supera gli ambiti dell’architettura sacra per riferirsi alla più profonda chiave interpretativa di architettura cultuale, concezione in genere sovente trascurata se non ignorata del tutto. Il loro operato rischiara infatt…
Nucleotide excision repair of abasic DNA lesions
2019
AbstractApurinic/apyrimidinic (AP) sites are a class of highly mutagenic and toxic DNA lesions arising in the genome from a number of exogenous and endogenous sources. Repair of AP lesions takes place predominantly by the base excision pathway (BER). However, among chemically heterogeneous AP lesions formed in DNA, some are resistant to the endonuclease APE1 and thus refractory to BER. Here, we employed two types of reporter constructs accommodating synthetic APE1-resistant AP lesions to investigate the auxiliary repair mechanisms in human cells. By combined analyses of recovery of the transcription rate and suppression of transcriptional mutagenesis at specifically positioned AP lesions, w…
Improvement of baculovirus as protein expression vector and as biopesticide by CRISPR/Cas9 editing
2019
The clustered regularly interspaced short palindromic repeats (CRISPR) system?associated Cas9 endonuclease is a molecular tool that enables specific sequence editing with high efficiency. In this study, we have explored the use of CRISPR/Cas9 system for the engineering of baculovirus. We have shown that the delivering of Cas9-single guide RNA ribonucleoprotein (RNP) complex with or without DNA repair template into Sf21 insect cells through lipofection might be efficient to produce knockouts as well as knock-ins into the baculovirus. To evaluate potential application of our CRISPR/Cas9 method to improve baculovirus as protein expression vector and as biopesticide, we attempted to knockout se…