pSPCas9(BB)-2A-GFP plasmids

CRISPR Mouse - Deletion 3T3-L1 Usp2

Experiment
CRISPR Mouse - Deletion 3T3-L1 Usp2
Product
pSPCas9(BB)-2A-GFP plasmids from Addgene
Manufacturer
Addgene

Protocol tips

Protocol tips
There is no need to perform a negative control golden-gate reaction (without
insert) as it will always contain colonies so not a good indicator of cloning success.

Publication protocol

2.1. Cells
Mouse peritoneal macrophages were prepared as described previously [14]. USP2 knockdown (KD) and their control cells were generated based on human myeloid HL-60 cells [13]. One day prior to examination, the cells were treated with phorbol 12-myristate 13-acetate (30 nM; Sigma-Aldrich, St Louis, MO, USA) to induce macrophage-like differentiation. Expression levels of Usp2 variants were downregulated by ~20% relative to control cells [13]. Mouse 3T3-L1 cells were purchased from Takara Bio (Kusatsu, Japan) and were differentiated into mature adipocytes by adding Adipoinducer cocktail for one week (Takara Bio). Mouse C2C12 myoblasts were obtained from RIKEN BioResource Center (Tsukuba, Japan), and were differentiated into myotubes in Dulbecco's modified Eagle medium (DMEM) (Nacalai, Kyoto, Japan) containing 4.5 g/L of glucose and 2% horse serum (Thermo Fischer Scientific, Waltham, MA, USA) for five days.

Full paper   Login or join for free to view the full paper.

Reviews

pSPCas9(BB)-2A-GFP plasmids from Addgene has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Discussion

4 years ago

Author: Mario Udinese Italy

Floxing mice with CRISPR

Hi everyone! I am planning on floxing mice with CRISPR but I am having trouble deciding which region to target. Do you have any tips on choosing?

Discussion

5 years ago

Author: Ben Saar Israel

How to choose a region to target for CRISPR

Hi everyone! I am planning on floxing mice with CRISPR but I am having trouble deciding which region to target. Do you have any tips on choosing?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing CRISPR Mouse - Deletion 3T3-L1 Usp2 using pSPCas9(BB)-2A-GFP plasmids from Addgene.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Addgene for pSPCas9(BB)-2A-GFP plasmids below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing CRISPR Mouse - Deletion 3T3-L1 Usp2 using pSPCas9(BB)-2A-GFP plasmids from Addgene. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms