Repositorium
Journal Article / 2017
Jacobs, Thomas B.; Zhang, Ning; Patel, Dhruv; Martin, Gregory B.
The high efficiency of clustered regularly interspaced short palindromic repeats (CRISPR)-mediated mutagenesis in plants enables the development of high-throughput mutagenesis strategies. By transforming pooled CRISPR libraries into tomato (Solanum lycopersicum), collections of mutant lines were generated with minimal transformation attempts and in a relatively short period of time. Identification of the targeted gene(s) was easily determined by sequencing the incorporated guide RNA(s) in the primary transgenic events. From a single transformation with a CRISPR library targeting the immunity-associated leucine-rich repeat subfamily XII genes, heritable mutations were recovered in 15 of the 54 genes targeted. To increase throughput, a second CRISPR library was made containing three guide RNAs per construct to target 18 putative transporter genes. This resulted in stable mutations in 15 of the 18 targeted genes, with some primary transgenic plants having as many as five mutated genes. Furthermore, the redundancy in this collection of plants allowed for the association of aberrant T0 phenotypes with the underlying targeted genes. Plants with mutations in a homolog of an Arabidopsis (Arabidopsis thaliana) boron efflux transporter displayed boron deficiency phenotypes. The strategy described here provides a technically simple yet high-throughput approach for generating a collection of lines with targeted mutations and should be applicable to any plant transformation system.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
1214 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 PDS |
Albino phenotype SDN1 |
Basic research Basic research |
1215 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc07g016010 |
No information SDN1 |
Basic research Basic research |
1216 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc06g006040 |
No information SDN1 |
Basic research Basic research |
1217 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc10g085110 |
No information SDN1 |
Basic research Basic research |
1218 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc06g006020 |
No information SDN1 |
Basic research Basic research |
1219 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc03g006100 |
No information SDN1 |
Basic research Basic research |
1220 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc08g075620 |
No information SDN1 |
Basic research Basic research |
1221 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc02g068880 |
No information SDN1 |
Basic research Basic research |
1222 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc04g014890 |
No information SDN1 |
Basic research Basic research |
1223 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc04g014650 |
No information SDN1 |
Basic research Basic research |
1224 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc06g005880 |
No information SDN1 |
Basic research Basic research |
1225 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc02g031790 |
No information SDN1 |
Basic research Basic research |
1226 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc08g075600 |
No information SDN1 |
Basic research Basic research |
1227 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc10g085120 |
No information SDN1 |
Basic research Basic research |
1228 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc04g015980 |
No information SDN1 |
Basic research Basic research |
1229 |
Martin, Gregory B. USA |
Solanum lycopersicum |
CRISPR/Cas9 Solyc02g070890 |
No information SDN1 |
Basic research Basic research |