Screening for functional circular RNAs using the CRISPR-Cas13 system
Summary Circular RNAs (circRNAs) produced from back-spliced exons are widely expressed, but individual circRNA functions remain poorly understood due to inadequate methods, such as RNAi and genome engineering, in distinguishing overlapped exons in circRNAs from those in linear cognate mRNAs1,2. Here we report that the programable RNA-guided, RNA-targeting CRISPR-Cas13, RfxCas13d, effectively and specifically discriminates circRNAs from mRNAs, using guide (g)RNAs targeting sequences spanning the back-splicing junction (BSJ) sites featured in RNA circles. Using a lentiviral library that targets sequences across BSJ sites of highly expressed human circRNAs, we show that a group of circRNAs are important for cell growth mostly in a cell-type specific manner and that a common oncogenic circRNA,circFAM120A,promotes cell proliferationin vitroandin vivoby preventingFAM120AmRNA from binding the translation inhibitor IGF2BP2 for efficient translation. Application of RfxCas13d/BSJ-gRNA screening has also uncoveredcircMan1a2with regulatory potential in mouse embryo preimplantation development. Together, these results establish CRISPR-RfxCas13d as a useful tool for the discovery and functional study of circRNAs at both individual and large-scale levels..
| Medienart: |
|
|---|
Preprint| Erscheinungsjahr: |
2021 |
|---|---|
| Erschienen: |
2021 |
| Enthalten in: |
bioRxiv.org - (2021) vom: 06. März Zur Gesamtaufnahme - year:2021 |
|---|
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Li, Siqi [VerfasserIn] |
|---|
| Links: |
Volltext [kostenfrei] |
|---|
| doi: |
10.1101/2020.03.23.002865 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
XBI000816159 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | XBI000816159 | ||
| 003 | DE-627 | ||
| 005 | 20230429090004.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 200327s2021 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1101/2020.03.23.002865 |2 doi | |
| 035 | |a (DE-627)XBI000816159 | ||
| 035 | |a (DE-599)biorXiv10.1101/2020.03.23.002865 | ||
| 035 | |a (biorXiv)10.1101/2020.03.23.002865 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 082 | 0 | |a 570 |q DE-84 | |
| 100 | 1 | |a Li, Siqi |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Screening for functional circular RNAs using the CRISPR-Cas13 system |
| 264 | 1 | |c 2021 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 520 | |a Summary Circular RNAs (circRNAs) produced from back-spliced exons are widely expressed, but individual circRNA functions remain poorly understood due to inadequate methods, such as RNAi and genome engineering, in distinguishing overlapped exons in circRNAs from those in linear cognate mRNAs1,2. Here we report that the programable RNA-guided, RNA-targeting CRISPR-Cas13, RfxCas13d, effectively and specifically discriminates circRNAs from mRNAs, using guide (g)RNAs targeting sequences spanning the back-splicing junction (BSJ) sites featured in RNA circles. Using a lentiviral library that targets sequences across BSJ sites of highly expressed human circRNAs, we show that a group of circRNAs are important for cell growth mostly in a cell-type specific manner and that a common oncogenic circRNA,circFAM120A,promotes cell proliferationin vitroandin vivoby preventingFAM120AmRNA from binding the translation inhibitor IGF2BP2 for efficient translation. Application of RfxCas13d/BSJ-gRNA screening has also uncoveredcircMan1a2with regulatory potential in mouse embryo preimplantation development. Together, these results establish CRISPR-RfxCas13d as a useful tool for the discovery and functional study of circRNAs at both individual and large-scale levels. | ||
| 700 | 1 | |a Li, Xiang |e verfasserin |4 aut | |
| 700 | 1 | |a Xue, Wei |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Lin |e verfasserin |4 aut | |
| 700 | 1 | |a Cao, Shi-Meng |e verfasserin |4 aut | |
| 700 | 1 | |a Lei, Yun-Ni |e verfasserin |4 aut | |
| 700 | 1 | |a Yang, Liang-Zhong |e verfasserin |4 aut | |
| 700 | 1 | |a Guo, Si-Kun |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Jia-Lin |e verfasserin |4 aut | |
| 700 | 1 | |a Gao, Xiang |e verfasserin |4 aut | |
| 700 | 1 | |a Wei, Jia |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Jinsong |e verfasserin |4 aut | |
| 700 | 1 | |a Yang, Li |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Ling-Ling |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t bioRxiv.org |g (2021) vom: 06. März |
| 773 | 1 | 8 | |g year:2021 |g day:06 |g month:03 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1101/2020.03.23.002865 |z kostenfrei |3 Volltext |
| 912 | |a GBV_XBI | ||
| 912 | |a SSG-OLC-PHA | ||
| 951 | |a AR | ||
| 952 | |j 2021 |b 06 |c 03 | ||
| 953 | |2 045F |a 570 | ||