Virally induced CRISPR/Cas9-based knock-in of fluorescent albumin allows long-term visualization of cerebral circulation in infant and adult mice
Vittani, M.; Knak, P. A. G.; Fukuda, M.; Nagao, M.; Wang, X.; Kjaerby, C.; Konno, A.; Hirai, H.; Nedergaard, M.; Hirase, H.
Show abstract
Albumin, a protein produced by liver hepatocytes, represents the most abundant protein in blood plasma. We have previously engineered a liver-targeting adeno-associated viral vector (AAV) that expresses fluorescent protein-tagged albumin to visualize blood plasma in mice. While this approach is versatile for imaging in adult mice, transgene expression vanishes when AAV is administered in neonates due to dilution of the episomal AAV genome in the rapidly growing liver. Here, we use CRISPR/Cas9 genome editing to insert the fluorescent protein mNeonGreen (mNG) gene into the albumin (Alb) locus of hepatocytes to produce fluorescently labeled albumin (Alb-mNG). We constructed a CRISPR AAV that includes [~]1 kb homologous arms around Alb exon 14 to express Alb-mNG. Subcutaneous injection of this AAV with AAV-CMV-Cas9 in postnatal day 3 mice resulted in two-photon visualization of the cerebral cortex vasculature within ten days. The expression levels of Alb-mNG were persistent for at least three months and were so robust that vasomotion and capillary blood flow could be assessed transcranially in early postnatal mice. This knock-in approach provides powerful means for micro- and macroscopic imaging of cerebral vascular dynamics in postnatal and adult mice.
Matching journals
The top 7 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Efficient Gene Transduction in Pigs and Macaques with the Engineered AAV Vector AAV.GT5 for Hemophilia B Gene Therapy 93%
- Selection of rAAV Vectors that Cross the Human Blood-Brain Barrier 1 and Target the Central Nervous System Using a Transwell Model 93%
- Small Alphaherpesvirus Latency-Associated Promoters Drive Efficient And Long-Term Transgene Expression In The Central Nervous System 92%
Similar papers in this journal
- Gene Therapy Prevents Hepatic Mitochondrial Dysfunction In Murine Deoxyguanosine Kinase Deficiency 94%
- Transcriptional changes in non-human primate tissues after intrathecal delivery of serotype 9 adeno-associated viral vector: insights into organ toxicities 94%
- A novel functional gene delivery platform based on a commensal human anellovirus demonstrates transduction in multiple tissue types 92%
Similar papers in this journal
- Enhancing gene transfer to renal tubules and podocytes by context-dependent selection of AAV capsids 94%
- In vivo lentiviral vector gene therapy to cure hereditary tyrosinemia type 1 and prevent development of precancerous and cancerous lesions 93%
- AAV11 permits efficient retrograde targeting of projection neurons 93%
Similar papers in this journal
- Administration of barcoded AAV capsid library to the putamen of non-human primates identifies variants with efficient retrograde transport 93%
- AAV-mediated gene therapy for Sialidosis 93%
- Preclinical lentiviral vector-mediated hematopoietic stem and progenitor cell gene therapy corrects Pompe disease-related muscle and neurological manifestations 93%
Similar papers in this journal
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.