Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

ABSTRACT: Cas12a ortholog cleavage pattern and PAM assay

PROVIDER: PRJNA631245 | ENA |

REPOSITORIES: ENA

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	SRR11748920_1.fastq.gz	Fastqsanger.gz
	SRR11748920_2.fastq.gz	Fastqsanger.gz
	SRR11748921_1.fastq.gz	Fastqsanger.gz
	SRR11748921_2.fastq.gz	Fastqsanger.gz
	SRR11748922_1.fastq.gz	Fastqsanger.gz

Items per page:

1 - 5 of 64

Similar Datasets

Randomized PAM depletion of phylogenetically-diverse Cas12a nucleases

Project description:We report the PAMs of phylogenetically-diverse Cas12a nucleases using a cell-free TXTL-based PAM screen. By adding a 5N randomized PAM library and Cas12a-gRNA in vitro, recognized PAM sequences were cleaved, while non-recognized PAMs remained. By amplifying the non-cleaved DNA, we used next-generation sequencing to analyze the depletion of functional PAMs of these Cas12a nucleases.

2020-03-02 | GSE130377 | GEO

The compact high-fidelity EbCas12a enables all-in-one AAV delivery system

Project description:we characterized a novel compact Cas12a ortholog, EbCas12a, from the Erysipelotrichia bacterium with activities in mammalian cells. It is with the PAM sequence of 5’-TTTV-3’ (V=A, G, C) and the smallest size of ~3.47kb among reported Cas12a orthologs so far. Moreover, enhanced EbCas12a (enEbCas12a) was also developed to have comparable editing efficiency with higher specificity to AsCas12a and LbCas12a in mammalian cells. With the help of the compact enEbCas12a, all-in-one AAV delivery system with crRNA for Cas12a was developed for both in vitro and in vivo. Altogether, with the help of the novel smallest high fidelity enEbCas12a, this first case of the all-in-one AAV delivery for Cas12a could greatly boost future gene therapy and scientific research.

2023-07-10 | GSE236647 | GEO

Randomized PAM depletion of AsCas12a

Project description:We report the PAMs of AsCas12a using a cell-free TXTL-based cleavage assay. By adding randomized PAM library and AsCas12a-gRNA in vitro, functional PAM sequences were cleaved, while non-functional PAMs remained. By amplifying the non-cleaved DNA, we use next-generation sequencing to analyze the depletion of functional PAMs of AsCas12a.

2019-01-01 | GSE123443 | GEO

Randomized PAM depeletion of NmeCas9

Project description:We report the PAMs of NmeCas9 using a cell-free TXTL-based cleavage assay. By adding randomized PAM library and NmeCas9-gRNA in vitro, functional PAM sequences were cleaved, while non-functional PAMs remained. By amplifying the non-cleaved DNA, we use next-generation sequencing to analyze the depletion of functional PAMs of NmeCas9.

2017-12-21 | GSE108357 | GEO

Homo sapiens

Project description:Novel Cas12a, broad PAM editor

| PRJNA1043844 | ENA

Project description:A new Cas12a with TTNA PAM and robust collateral cleavage for nucleic acid detection

| PRJNA783665 | ENA

Detecting non-canonically-targeted SpCas9 generated DSBs in human cells by LAM-HTGTS

Project description:We identified robust cleavage by non-canonically-targeted SpCas9 that target protospacer with a few bases from the canonical NGG PAM in biochemistry assay. To determine whether this spaced SpCas9 cleavage exists in human cells, we employed high-throughput genome-wide translocation sequencing (LAM-HTGTS) using SpCas9 with three independent canonical sgRNAs served as bait (bait-A, bait-L and bait-D) to compared the cleavage activities of canonically-targeted and non-canonically-targeted SpCas9 in eight locus (RAG1B-D, RAG1K-O) in chromosome 11. We found that almost all 1bp-target shifted SpCas9 generated substantial DSBs as the canonical controls. While detected less frequently, 2bp-targeted shifted SpCas9 generated significant amounts of DSBs in RAG1D and RAG1L locus. These findings underscore the spaced PAM-mediated DSBs occurred in living cells.

2022-02-28 | GSE192459 | GEO

synthetic construct

Project description:CjCas9 PAM discovery assay

| PRJNA993711 | ENA

A Cas12a ortholog with stringent PAM recognition followed by low off-target editing rates for genome editing

Project description:A Cas12a ortholog with stringent PAM recognition followed by low off-target editing rates for genome editing

| PRJNA610555 | ENA

synthetic metagenome

Project description:Cas9 ortholog cut site assay

| PRJNA622541 | ENA