| RRC ID |
36142
|
| Author |
Zhu L, Mon H, Xu J, Lee JM, Kusakabe T.
|
| Title |
CRISPR/Cas9-mediated knockout of factors in non-homologous end joining pathway enhances gene targeting in silkworm cells.
|
| Journal |
Sci Rep
|
| Abstract |
Gene targeting can be achieved by precise genetic modifications through homology-directed repair (HDR) after DNA breaks introduced by genome editing tools such as CRISPR/Cas9 system. The most common form of HDR is homologous recombination (HR). Binding to the DNA breaks by HR factors is thought to compete with non-homologous end joining (NHEJ), an alternative DNA repair pathway. Here, we knocked out the factors in NHEJ by CRISPR/Cas9 system in silkworm cells, so that increased the activities of HR up to 7-fold. Also efficient HR-mediated genome editing events occurred between the chromosomal BmTUDOR-SN gene and donor DNA sequences with an EGFP gene in the middle of two homologous arms for the target gene. Utilizing the NHEJ-deficient silkworm cells, we found that homologous arms as short as 100 bp in donor DNA could be designed to perform precise genome editing. These studies should greatly accelerate investigations into genome editing of silkworm.
|
| Volume |
5
|
| Pages |
18103
|
| Published |
2015-12-10
|
| DOI |
10.1038/srep18103
|
| PII |
srep18103
|
| PMID |
26657947
|
| PMC |
PMC4674802
|
| MeSH |
Animals
Animals, Genetically Modified
Base Sequence
Bombyx / cytology
Bombyx / genetics*
CRISPR-Cas Systems*
Cells, Cultured
DNA End-Joining Repair*
Gene Knockout Techniques / methods*
Gene Targeting / methods*
Genetic Engineering / methods
Genome, Insect / genetics
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
Homologous Recombination
Insect Proteins / genetics
Insect Proteins / metabolism
Microscopy, Fluorescence
Molecular Sequence Data
Signal Transduction / genetics
|
| IF |
3.998
|
| Times Cited |
15
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Silkworms |
|
