WebOct 10, 2024 · This study uses a genome-wide CRISPR-Cas9 screen to reveal new factors that influence the accumulation of Replication Protein A on DNA after UV irradiation; ... Schematic overview of the FACS-based CRISPR-Cas9 screen. Cells were irradiated with 1 J/m 2 UV at 6, 9, 12, and 15 days postinfection with the GeCKOv2 lentiviral library ... WebCas9 screen技术是一项极具潜力的高通量筛选工具，已被应用于各个领域。 同时，其衍生与拓展技术，如Cas13 screen、Base editor screen也逐步问世。 我相信未来的研究手 …
Genome-wide CRISPR screens identify ILF3 as a mediator …
THP-1 iCas9 cells were transduced with lentiviral particles using spinfection. To this end, THP-1 cells were transferred into 6-well plates, virus was added and plates were centrifuged at 1000×g for 20 min. Two days (genome-wide screen) or five days later (validation screen), transduced cells were MACS enriched using … See more THP-1 (TIB-202 from ATCC) cells were cultured in RPMI1640 + Glutamax-I Medium (Gibco, 61870-010) with 10% FCS (Invitrogen, … See more Design, construction, and basic performance of the genome-wide sgRNA library has been described in Michlits et al.21. To generate focused sgRNA pools for validation screens, oligo pools were obtained from Twist … See more FACS-enriched or baseline cells were collected by centrifugation, washed with PBS and a maximum of 5E6 cells were resuspended in 350 µL DNA extraction buffer (10 mM Tris-HCl, 150 mM NaCl, 10 m M EDTA, pH … See more Lentiviral vectors were produced using MISSION Lentiviral Packaging Mix (Sigma-Aldrich), PEI Max (Polysciences, Inc.) and Lenti X-293T cells (Clontech Laboratories, Inc) according to the manufacturer’s … See more WebNov 14, 2024 · We call this technique FACS-assisted CRISPR-Cas9 editing (FACE). FACE allows the derivation of correctly edited polyclones carrying a positive selection fluorescent module and the exclusion of non-edited, random integrations and on-target allele NHEJ-containing cells. We derived a set of isogenic lines containing Parkinson's-disease … kmcc airport
A Genome-wide CRISPR Screen Identifies WDFY3 as a Novel
WebNov 6, 2024 · CRISPR screening is a large-scale genetic loss-of-function experimental approach designed to find the equivalent of a few needles in a haystack. CRISPR screening facilitates discovery of key genes or … Web• Developed an assay by using CRISPR/Cas9 mediated gene-disruption combined with high-throughput flow cytometry-based competition … WebJan 22, 2024 · Phagocytic clearance of dying cells, termed efferocytosis, must occur efficiently to maintain homeostasis and prevent disease. Yet, our understanding of this important biological process remains incomplete. To search for novel regulators of efferocytosis, we performed a FACS-based genome-wide CRISPR knockout screen in … red band not rated