Reference - Detail
| RRC ID | 74622 |
|---|---|
| Author | Mehta S, Buyanbat A, Orkin S, Nabet B. |
| Title | High-efficiency knock-in of degradable tags (dTAG) at endogenous loci in cell lines. |
| Journal | Methods Enzymol |
| Abstract |
The dTAG system is a versatile strategy for tunable control of protein abundance and facilitates the time-resolved assessment of disease-associated protein function. A "co-opted" fusion-based degron peptide, the "dTAG" facilitates the study of endogenous protein function when knocked-in at the endogenous genetic loci of proteins of interest. We combine CRISPR/Cas9 mediated induction of double-strand breaks (DSB) with the delivery of a single-stranded DNA HDR-donor-template via crude preparations of recombinant adeno-associated virus (rAAV). Our approach to knock-in of large (1-2kb) DNA fragments via crude-rAAV mediated HDR donor delivery is rapid and inexpensive. It facilitates genetic modification of a variety of human as well as mouse cell lines at high efficiency and precision. |
| Volume | 681 |
| Pages | 1-22 |
| Published | 2023-1-1 |
| DOI | 10.1016/bs.mie.2022.08.045 |
| PII | S0076-6879(22)00370-6 |
| PMID | 36764753 |
| MeSH | Animals CRISPR-Cas Systems* DNA DNA, Single-Stranded Gene Editing* Humans Mice Recombinational DNA Repair |
| Altmetric score |
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 27 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | HUDEP-2(RCB4557) |