| RRC ID |
51484
|
| Author |
Prior H, Jawad AK, MacConnachie L, Beg AA.
|
| Title |
Highly Efficient, Rapid and Co-CRISPR-Independent Genome Editing in Caenorhabditis elegans.
|
| Journal |
G3 (Bethesda)
|
| Abstract |
We describe a rapid and highly efficient method to generate point mutations in Caenorhabditis elegans using direct injection of CRISPR-Cas9 ribonucleoproteins. This versatile method does not require sensitized genetic backgrounds or co-CRISPR selection-based methods, and represents a single strategy that can be used for creating genomic point mutations, regardless of location. As proof of principle, we show that knock-in mutants more faithfully report variant-associated phenotypes as compared to transgenic overexpression. Data for nine knock-in mutants across five genes are presented that demonstrate high editing efficiencies (60%), a reduced screening workload (24 F1 progeny), and a rapid timescale (4-5 d). This optimized method simplifies genome engineering and is readily adaptable to other model systems.
|
| Volume |
7(11)
|
| Pages |
3693-3698
|
| Published |
2017-11-6
|
| DOI |
10.1534/g3.117.300216
|
| PII |
g3.117.300216
|
| PMID |
28893845
|
| PMC |
PMC5677160
|
| MeSH |
Animals
CRISPR-Cas Systems*
Caenorhabditis elegans / genetics*
Gene Editing / methods*
Gene Knockout Techniques / methods*
Genome, Helminth
Point Mutation
|
| IF |
2.781
|
| Times Cited |
12
|
| Resource |
| C.elegans |
tm1444 |
