crispr-human-deletion-f11r

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

Get tips on using pX333-sgNF1-sgTrp53-Cas9 to perform CRISPR Mouse - Deletion 3T3-L1 Trp53

Get tips on using pSpCas9(BB)-2A-GFP (PX458) to perform CRISPR Rat - Deletion AR42J FICD

Get tips on using pSpCas9(BB)-2A-GFP (PX458) to perform CRISPR Rat - Deletion AR42J Atg12

Get tips on using pX330-U6-Chimeric_BB-CBh-hSpCas9 to perform CRISPR Rat - Deletion BMSCs Wisp2

Get tips on using pSpCas9(BB)-2A-GFP (PX458) to perform CRISPR Rat - Deletion PC12 Munc18

Get tips on using pSpCas9(BB)-2A-GFP (PX458) to perform CRISPR Mouse - Deletion SL4 Ptgs2

Get tips on using pSpCas9(BB)-2A-Puro (PX459) to perform CRISPR Mouse - Deletion L929 SIRT2

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.