Costruzione di plasmidi CRISPR e rilevamento dell'efficienza knockout nelle cellule di mammifero attraverso un sistema Dual Luciferase Reporter
6,244 Views
•
05:51 min
•
December 05, 2020
DOI:
Summary
Automatically generated
Qui presentiamo un protocollo che descrive un metodo semplificato per la generazione efficiente di plasmidi che esprimono sia l'enzima CRISPR che l'RNA a guida singola associato (sgRNA). La co-trasfezione delle cellule di mammifero con questo vettore sgRNA/CRISPR e un vettore reporter a doppia luciferasi che esamina la riparazione della rottura a doppio filamento consente la valutazione dell'efficienza knockout.
Video correlati
08:25
Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format
Video correlati
13850 Views
11:35
Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
Video correlati
12499 Views
07:56
Genome Editing in Mammalian Cell Lines using CRISPR-Cas
Video correlati
21581 Views
00:09
Pooled CRISPR-Based Genetic Screens in Mammalian Cells
Video correlati
21771 Views
08:49
Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
Video correlati
14487 Views
10:13
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
Video correlati
11090 Views
10:38
Monitoring Cell-autonomous Circadian Clock Rhythms of Gene Expression Using Luciferase Bioluminescence Reporters
Video correlati
22352 Views
12:34
Monitoring Plasmid Replication in Live Mammalian Cells over Multiple Generations by Fluorescence Microscopy
Video correlati
11792 Views
12:55
High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay
Video correlati
23538 Views
11:32
Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines
Video correlati
10445 Views
Read Article
Cite this Article
Li, H., Qin, H., Zhang, N., Zhao, J., Xin, J., Perez-Campo, F. M., Liu, H. Construction of CRISPR Plasmids and Detection of Knockout Efficiency in Mammalian Cells through a Dual Luciferase Reporter System. J. Vis. Exp. (166), e59639, doi:10.3791/59639 (2020).
Copy