Year: 2023
Type: article
Abstract: Microinjected transgenes, both large and small, are known to insert randomly into the mouse genome. Traditional methods of mapping a transgene are challenging, thus complicating breeding strategies and accurate interpretation of phenotypes, particularly when a transgene disrupts critical coding or noncoding sequences. As the vast majority of transgenic mouse lines remain unmapped, we developed CRISPR-Cas9 Long-Read Sequencing (CRISPR-LRS) to ascertain transgene integration loci. This novel approach mapped a wide size range of transgenes and uncovered more complex transgene-induced host genome re-arrangements than previously appreciated. CRISPR-LRS offers a facile, informative approach to establish robust breeding practices and will enable researchers to study a gene without confounding genetic issues. Finally, CRISPR-LRS will find utility in rapidly and accurately interrogating gene/genome editing fidelity in experimental and clinical settings.
Source: The CRISPR Journal
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: Novel Interplay of KILN and MKL1 in Vascular Pathophysiology, Transcriptional Control of Myocardin and the MYOCARDome, G4681031153, Function and Regulation of TSPAN2 in Vascular Disease, G6324451732, Role of VAMP1 in synaptic transmission and Alzheimer's dementia, Regulation and Function of SRF in Vascular Pathiobiology, Role of cyclic nucleotide signaling in aortic aneurysm
OpenAlex ID: W4366235011