Publications by authors named "Amr R Salem"
Article Synopsis
- Traditional methods of genetically modifying mice are complex and time-consuming, requiring advanced techniques and often taking over a year with uncertain outcomes.
- The CRISPR gene-editing system has simplified this process, allowing for efficient and successful modifications in mice and other animal models.
- This text introduces a streamlined approach using prime editing, detailing three basic protocols for creating genetically modified mice: designing engineered RNA, microinjecting components into zygotes, and genotyping founder mice for breeding.
CRISPR editing involves double-strand breaks in DNA with attending insertions/deletions (indels) that may result in embryonic lethality in mice. The prime editing (PE) platform uses a prime editing guide RNA (pegRNA) and a Cas9 nickase fused to a modified reverse transcriptase to precisely introduce nucleotide substitutions or small indels without the unintended editing associated with DNA double-strand breaks. Recently, engineered pegRNAs (epegRNAs), with a 3'-extension that shields the primer-binding site of the pegRNA from nucleolytic attack, demonstrated superior activity over conventional pegRNAs in cultured cells.
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