...targeting the region where the edit should be made HDR requires a repair template and displays reduced efficiency...protein domains. For gene editing experiments using HDR, it is essential that the target sequence be very...different owing to the error-prone nature of NHEJ. In HDR gene editing experiments, most mutated alleles will...identify DNA fragments. PCR and restriction digest (for HDR repaired DSBs): For small nucleotide edits that introduce.... PCR amplification and gel electrophoresis (for HDR or NHEJ): For large deletions or insertions, the ...amplification, subcloning and Sanger sequencing (for HDR or NHEJ): Provides semi-quantitative assessment of...amplification and next-generation sequencing (for HDR or NHEJ): Provides quantitative assessment of the...

...Modifications Using Homology Directed Repair (HDR) Figure 4: Overview of HDR HDR generates specific genomic edits ...but high-fidelity homology directed repair (HDR) pathway. HDR relies on copying DNA from a matching template... Both NHEJ and HDR have their uses, but we’ll focus on NHEJ first and come back to HDR . NHEJ is a naturally...that modulate HDR or NHEJ components, like miCas9 , Cas9-DN1S , or others, can increase HDR efficiency. ...base. These small precision edits can be done via HDR, base editing, and prime editing, and are often used...mutation that does not change the amino acid sequence. HDR efficiency is generally low (< 10% of modified alleles...Synchronizing cells or modulating the cell cycle can help as HDR takes place during the S and G2 phases. Chemically...