Generation of Stable Expression Mammalian Cell Lines Using Lentivirus
[Abstract] Lentiviruses are used very widely to generate stable expression mammalian cell lines. They are used for both gene down-regulation (by using shRNA) or for gene up-regulation (by using ORF of gene of interest). The technique of generating stable cell lines using 3rd generation lentivirus is very robust and it typically takes about 1-2 ...
Qualitative in vivo Bioluminescence Imaging
[Abstract] Bioluminescence imaging (BLI) technology is an advanced method of carrying out molecular imaging on live laboratory animals in vivo. This powerful technique is widely-used in studying a variety of biological processes, and it has been an ideal tool in exploring tumor growth and metastatic spread in real-time. This technique ensures the ...
Method for CRISPR/Cas9 Mutagenesis in Candida albicans
Authors: Neta Dean and
Henry Ng,
date: 04/20/2018,
view: 3714,
Q&A: 0
[Abstract] Candida albicans is the most prevalent and important human fungal pathogen. The advent of CRISPR as a means of gene editing has greatly facilitated genetic analysis in C. albicans. Here, we describe a detailed step-by-step procedure to construct and analyze C. albicans deletion mutants. This protocol uses plasmids that ...
Medaka-microinjection with an Upright Microscope
Authors: Yu Murakami and
Masato Kinoshita,
date: 02/05/2018,
view: 2349,
Q&A: 0
[Abstract] We described a simple method for microinjecting DNA/RNA/Protein solutions into medaka eggs under an upright microscope. Medaka is an excellent vertebrate model for reverse genetics, because of its daily spawning, short generation time, and egg transparency. These features enable us to efficiently perform functional genomic analyses of transgenic ...
Generation of Chemically Induced Liver Progenitors (CLiPs) from Rat Adult Hepatocytes
[Abstract] Primary mature hepatocytes (MHs) or their progenitor cells are candidate cell sources for cell transplantation therapy in severe liver diseases. However, stable culture of these cells or generation of equivalent cells from pluripotent stem cells has been limited. Using a cocktail of small molecules that we previously found useful in stable culture ...
Ex vivo Trophoblast-specific Genetic Manipulation Using Lentiviral Delivery
[Abstract] In this protocol report, we describe a lentiviral gene delivery technique for genetic modification of the rat trophoblast cell lineage. Lentiviral packaged gene constructs can be efficiently and specifically delivered to the trophoblast cell lineage of the blastocyst. The consequences of ‘gain-of-function’ and ‘loss-of-function’ blastocyst ...