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Credit: CC0 Public Domain
Scientists at Wake Forest's Institute for Regenerative Medicine (WFIRM) have discovered a better way to provide a tool for editing DNA to reduce the presence of editing proteins in cells in what they describe as a random approach.
CRISPR technology (clustered regularly spaced short palindromic repeats) is used to modify DNA sequences and modify gene function. CRISPR / Cas9 is an enzyme used as a pair of scissors to cut two strands of DNA at a specific location to add, delete or repair DNA fragments. But CRISPR / Cas9 is not 100% accurate and could potentially cut unexpected locations, resulting in unwanted results.
"One of the major challenges of CRISPR / Cas9 mRNA technologies is the possibility of noncompliant targets that can cause tumors or mutations," said Baisong Lu, Ph.D., assistant professor of regenerative medicine at WFIRM and one of the main authors of the project. paper. While other types of lentivirus-like bionanoparticles (LVLPs) have been described to deliver proteins or mRNAs, Dr. Lu said, "The LVLP we have developed has unique features that will make it a useful tool. in the toolbox being edited the expanding genome ".
To solve the problem of inaccuracy, WFIRM researchers asked the following question: is there a way to efficiently deliver Cas9 activity while ensuring transient protein expression? of editing the genome? They tested various strategies and then took the best properties of two widely used vectors – the lentivirus vector and the nanoparticles – and combined them to create a system that effectively encapsulates Cas9 mRNA in LVLPs, allowing transient expression and a very effective edition.
The lentiviral vector is a gene transmission vehicle widely used in research laboratories and is already widely used to deliver CRISPR / Cas9 mRNA technology for efficient genome editing. Nanoparticles are also used but they are not as effective in delivering CRISPR / Cas9.
The WFIRM team published its findings in an article recently published in the journal Nucleic acid research.
"By combining the transient expression characteristic of nanoparticle delivery strategies while retaining the efficiency of lentiviral vector transduction, we have created a system that can be used to condition various protein mRNAs." edition for the purpose of genome editing, "randomly", "Anthony Atala, MD, director of WFIRM and co-lead author of the document. "This system will not only improve safety, but also avoid a possible immune response to the editor's proteins, which could improve the efficiency of gene editing in vivo, which will be useful in research and clinical applications. "
Explore further:
A new strategy improves the efficiency of CRISPR-Cas9 genome editing
More information:
Baisong Lu et al, Deletion of SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing Nucleic acid research (2019). DOI: 10.1093 / n / a / gkz093
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