CRISPR Mouse Deletion ES (embryonic stem) cells

Get tips on using hCas9 to perform CRISPR Mouse - Deletion 3T3-L1 ATP7A

Get tips on using lentiCRISPR v2 to perform CRISPR Mouse - Deletion NSC34 PGRMC1

Get tips on using lentiCRISPR v2 to perform CRISPR Mouse - Deletion ATDC5 Nfatc1

Get tips on using pX330A-1x3 to perform CRISPR Mouse - Deletion ATDC5 Perk

Get tips on using gRNA_Cloning Vector to perform CRISPR Mouse - Deletion L929 Gβ2

Get tips on using pSpCas9 (PX165) to perform CRISPR Mouse - Deletion C2C12 Klf5

Get tips on using pSpCas9 (PX165) to perform CRISPR Mouse - Deletion αT3 IP3R1

Get tips on using JAM-A CRISPR/Cas9 KO Plasmid (h) to perform CRISPR Human - Deletion F11R

Human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) have been greatly used for studies on embryonic development and cell differentiation.iPSCs provide a stable source for either self-renewal or differentiation into suitable cells when cultured in a particular environment. Pluripotent cell culture was originally started by deriving cells from inner cell mass (ICM) from pre-implanted blastocysts, these were called embryonic stem cells. These cells after isolation can be grown on traditional extracellular matrices (like mouse embryonic fibroblasts, MEFs) or feeder-free culture systems. DMEM/F12 has been the most commonly used basal media in the culture of pluripotent cells. These cells are cultured at normal atmospheric oxygen levels, 21%, however, some studies have proposed that 4% oxygen tension may be better for hESC growth. Higher D-glucose concentration (4.2g/l) and osmolarity (320mOsm) that mimics the natural environment of embryonic tissue are optimal for the growth of hESCs. Supplements like N2 and/or B-27, in the presence of growth factors like bFGF, have been shown to increase pluripotency of these cells. bFGF, FGF2 and other ligands of receptor tyrosine kinases like IGF are also required or maintain self-renewal ability of these cells. TGF𝛃1, by its activation of SMAD2/3 signalling, also represses differentiation of iPSCs. Other compounds like ROCK inhibitors reduce blebbing and apoptosis in these cells to maintain their clonogenicity. However, an inhibitor for LIF (leukaemia inhibitory factor, which is one of the pluripotent genes) has an opposing effect. Therefore, it is important to understand the culture conditions and media composition that affect downstream signalling in hESCs or iPSCs that may lead to their differentiation.

Get tips on using CAG-Cas9 to perform CRISPR Mouse - Deletion Neuro 2a Rab38