pSPCas9(BB)-2A-GFP plasmids

Protocol tips

Upstream tips	Protocol tips	Downstream 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.

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.

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Reviews

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.

Manufacturer protocol

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

Download manufacturer protocol

Videos

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