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New Genomic Research Study Findings Reported from Hanyang University (Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein...

July 1, 2014



New Genomic Research Study Findings Reported from Hanyang University (Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA)

By a News Reporter-Staff News Editor at Life Science Weekly -- Researchers detail new data in Life Science Research. According to news originating from Seoul, South Korea, by NewsRx correspondents, research stated, "RNA-guided endonucleases (RGENs) derived from the CRISPR/Cas system represent an efficient tool for genome editing. RGENs consist of two components: Cas9 protein and guide RNA."

Our news journalists obtained a quote from the research from Hanyang University, "Plasmid-mediated delivery of these components into cells can result in uncontrolled integration of the plasmid sequence into the host genome, and unwanted immune responses and potential safety problems that can be caused by the bacterial sequences. Furthermore, this delivery method requires transfection tools. Here we show that simple treatment with cell-penetrating peptide (CPP)-conjugated recombinant Cas9 protein and CPP-complexed guide RNAs leads to endogenous gene disruptions in human cell lines. The Cas9 protein was conjugated to CPP via a thioether bond, whereas the guide RNA was complexed with CPP, forming condensed, positively charged nanoparticles. Simultaneous and sequential treatment of human cells, including embryonic stem cells, dermal fibroblasts, HEK293T cells, HeLa cells, and embryonic carcinoma cells, with the modified Cas9 and guide RNA, leads to efficient gene disruptions with reduced off-target mutations relative to plasmid transfections, resulting in the generation of clones containing RGEN-induced mutations. Our CPP-mediated RGEN delivery process provides a plasmid-free and additional transfection reagent-free method to use this tool with reduced off-target effects."

According to the news editors, the research concluded: "We envision that our method will facilitate RGEN-directed genome editing."

For more information on this research see: Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA. Genome Research, 2014;24(6):1020-1027. Genome Research can be contacted at: Cold Spring Harbor Lab Press, Publications Dept, 1 Bungtown Rd, Cold Spring Harbor, NY 11724, USA (see also Life Science Research).

The news correspondents report that additional information may be obtained from S. Ramakrishna, Hanyang University, Inst Bioengn & Biopharmaceut Res, Seoul 133791, South Korea. Additional authors for this research include A.K. Dad, J. Beloor, R. Gopalappa, S.K. Lee and H. Kim.

Keywords for this news article include: Asia, Seoul, South Korea, Life Science Research

Our reports deliver fact-based news of research and discoveries from around the world. Copyright 2014, NewsRx LLC


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Source: Life Science Weekly


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