Using transient inhibition of DNA mismatch repair during a permissive stage of development, we demonstrate highly efficient prime editing of mouse embryos with few unwanted, local byproducts (average 58% precise edit frequency, 0.5% on-target error frequency across 13 substitution edits at 8 sites), enabling same-generation phenotyping of founders.
Using transient inhibition of DNA mismatch repair during a permissive stage of development, we demonstrate highly efficient prime editing of mouse embryos with few unwanted, local byproducts , enabling same-generation phenotyping of founders. Whole-genome sequencing reveals that mismatch repair inhibition increases off-target indels at low-complexity regions in the genome without any obvious phenotype in mice.
To confirm that the observed increase in indels was the result of mMLH1dn, we repeated family-based WGS analysis in a pedigree of mice in which select progeny were microinjected at the two-cell stage with mMLH1dn mRNA and the+ 5 G > A pegRNA but without any editor , as well as a pedigree in which select progeny were microinjected with PEmax mRNA and the), but two of three mMLH1dn-injected embryos recapitulated the strong increase in −1 bp deletions near mononucleotide tracts observed in the...
Intriguingly, for several edits, we observed higher frequencies of precise editing in embryos than typical with transient delivery of mMLH1dn in cultured cells. Although several factors that differed between studies could explain this observation, cells may also be particularly amenable to prime editing in early development.
Following PCR1, 2 μl PCR1 product was transferred to a second PCR reaction to add flanking regions that included a 4-bp custom R1 index using 10 pmol each primer with 1x NEBNext Ultra II Q5 master mix with cycle conditions: 98 °C for 30 s, 8 cycles of 98 °C for 10 s, 64 °C for 20 s and 72 °C for 20 s, followed by 72 °C for 2 min.
United States Latest News, United States Headlines
Similar News:You can also read news stories similar to this one that we have collected from other news sources.
DNA and Chintai Partner to Launch DNA Deal Desk, A Fully On-Chain Investment PlatformCrypto Blog
Read more »
Honolulu hopes to unravel mysteries of long-unclaimed bodies using advanced DNA testingForensic pathologists hope advanced DNA testing technology will enable them to attach names to all the unidentified people at the Honolulu Medical Examiner's Office.
Read more »
Detailed answers from water well company: What are they using, not using to deter brown waterKSAT sent a list of questions to a water well company about how it’s trying to keep the water clean, and Tuesday the company sent detailed answers.
Read more »
Using a Galaxy S25 will be like using a more practical version of iPhone 16Martin, a tech journalist at PhoneArena, brings a unique blend of humor and insight to his work. His fascination with smartphones began with a Galaxy Young and evolved through a series of trades and upgrades, making him a self-proclaimed smartphone nerd.
Read more »
Mobile County Public Schools using AI to catch students using it to cheatMobile County Public Schools is using artificial intelligence to catch students using AI to plagiarize. This week, the school board approved a contract with Tur
Read more »
Pre-lit LED Christmas Trees: A More Energy-Efficient ChoicePre-lit Christmas trees with LED lights are a more energy-saving option compared to traditional string lights. Despite challenges, such as artificial trees often being made from PVC or polyethylene plastics imported from China or South Korea, there are some options made from recycled materials, making them a more sustainable choice.
Read more »