CR45722_CR45723_CR43839
Species: Drosophila melanogaster
Position: chr2L: 19469125-19471627
Known as:
Transcript: CR45722_CR45723_CR43839
Sequence: Download
Description:
We identified 128 testis-specific Drosophila lncRNAs and knocked out 105 of them using an optimized three-component CRISPR/Cas9 system. Among the lncRNA knockouts, 33 (31%) exhibited a partial or complete loss of male fertility, accompanied by visual developmental defects in late spermatogenesis. In addition, six knockouts were fully or partially rescued by transgenes in a trans configuration, indicating that those lncRNAs primarily work in trans. Furthermore, gene expression profiles for five lncRNA mutants revealed that testis-specific lncRNAs regulate global gene expression, orchestrating late male germ cell differentiation.
sgRNAs
| sgRNA_ID | Sequence | Position (Chr) | Position (Lnc) | Length | PAM | Type | Validity | Cell line | Note | Ref. |
|---|---|---|---|---|---|---|---|---|---|---|
| gRNA1 | GGGCAGACACGAGCAAAGTCC | 19469398-19469418(+) | gene body (near 5') | 21 | CGG | CRISPRko | Homologous recombination efficiency: 2.17% | embryo | Knockout phenotype: male fertility decreased by 30%-60% | [1] |
| gRNA2 | GGATTGGGATTGCGGGCAG | 19469465-19469483(+) | gene body (near 5') | 19 | CGG | CRISPRko | Homologous recombination efficiency: 2.17% | embryo | Knockout phenotype: male fertility decreased by 30%-60% | [1] |
| gRNA3 | GGAGTCGGACTCGGAATCCTT | 19471351-19471371(+) | gene body (near 3') | 21 | CGG | CRISPRko | Homologous recombination efficiency: 2.17% | embryo | Knockout phenotype: male fertility decreased by 30%-60% | [1] |
| gRNA4 | GGAAACTTGGAGCATTGGAG | 19471489-19471508(-) | gene body (near 3') | 20 | CGG | CRISPRko | Homologous recombination efficiency: 2.17% | embryo | Knockout phenotype: male fertility decreased by 30%-60% | [1] |
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Reference
1. Wen K, Yang L, Xiong T, Di C, Ma D, et al. (2016). Critical roles of long noncoding RNAs in Drosophila spermatogenesis. Genome Res 26(9): 1233-44.
