Pinned Repositories
A-counterdown-timer
设置事件名称和发生时间等参数，无边框置顶在特定位置
alist
阿里云盘列表
aliyundrive-webdav
阿里云盘 WebDAV 服务
Anti-Screenshot
一个强制截屏的xposed module
Assemblies-of-putative-SARS-CoV2-spike-encoding-mRNA-sequences-for-vaccines-BNT-162b2-and-mRNA-1273
RNA vaccines have become a key tool in moving forward through the challenges raised both in the current pandemic and in numerous other public health and medical challenges. With the rollout of vaccines for COVID-19, these synthetic mRNAs have become broadly distributed RNA species in numerous human populations. Despite their ubiquity, sequences are not always available for such RNAs. Standard methods facilitate such sequencing. In this note, we provide experimental sequence information for the RNA components of the initial Moderna (https://pubmed.ncbi.nlm.nih.gov/32756549/) and Pfizer/BioNTech (https://pubmed.ncbi.nlm.nih.gov/33301246/) COVID-19 vaccines, allowing a working assembly of the former and a confirmation of previously reported sequence information for the latter RNA. Sharing of sequence information for broadly used therapeutics has the benefit of allowing any researchers or clinicians using sequencing approaches to rapidly identify such sequences as therapeutic-derived rather than host or infectious in origin. For this work, RNAs were obtained as discards from the small portions of vaccine doses that remained in vials after immunization; such portions would have been required to be otherwise discarded and were analyzed under FDA authorization for research use. To obtain the small amounts of RNA needed for characterization, vaccine remnants were phenol-chloroform extracted using TRIzol Reagent (Invitrogen), with intactness assessed by Agilent 2100 Bioanalyzer before and after extraction. Although our analysis mainly focused on RNAs obtained as soon as possible following discard, we also analyzed samples which had been refrigerated (~4 ℃) for up to 42 days with and without the addition of EDTA. Interestingly a substantial fraction of the RNA remained intact in these preparations. We note that the formulation of the vaccines includes numerous key chemical components which are quite possibly unstable under these conditions-- so these data certainly do not suggest that the vaccine as a biological agent is stable. But it is of interest that chemical stability of RNA itself is not sufficient to preclude eventual development of vaccines with a much less involved cold-chain storage and transportation. For further analysis, the initial RNAs were fragmented by heating to 94℃, primed with a random hexamer-tailed adaptor, amplified through a template-switch protocol (Takara SMARTerer Stranded RNA-seq kit), and sequenced using a MiSeq instrument (Illumina) with paired end 78-per end sequencing. As a reference material in specific assays, we included RNA of known concentration and sequence (from bacteriophage MS2). From these data, we obtained partial information on strandedness and a set of segments that could be used for assembly. This was particularly useful for the Moderna vaccine, for which the original vaccine RNA sequence was not available at the time our study was carried out. Contigs encoding full-length spikes were assembled from the Moderna and Pfizer datasets. The Pfizer/BioNTech data [Figure 1] verified the reported sequence for that vaccine (https://berthub.eu/articles/posts/reverse-engineering-source-code-of-the-biontech-pfizer-vaccine/), while the Moderna sequence [Figure 2] could not be checked against a published reference. RNA preparations lacking dsRNA are desirable in generating vaccine formulations as these will minimize an otherwise dramatic biological (and nonspecific) response that vertebrates have to double stranded character in RNA (https://www.nature.com/articles/nrd.2017.243). In the sequence data that we analyzed, we found that the vast majority of reads were from the expected sense strand. In addition, the minority of antisense reads appeared different from sense reads in lacking the characteristic extensions expected from the template switching protocol. Examining only the reads with an evident template switch (as an indicator for strand-of-origin), we observed that both vaccines overwhelmingly yielded sense reads (>99.99%). Independent sequencing assays and other experimental measurements are ongoing and will be needed to determine whether this template-switched sense read fraction in the SmarterSeq protocol indeed represents the actual dsRNA content in the original material. This work provides an initial assessment of two RNAs that are now a part of the human ecosystem and that are likely to appear in numerous other high throughput RNA-seq studies in which a fraction of the individuals may have previously been vaccinated. ProtoAcknowledgements: Thanks to our colleagues for help and suggestions (Nimit Jain, Emily Greenwald, Lamia Wahba, William Wang, Amisha Kumar, Sameer Sundrani, David Lipman, Bijoyita Roy). Figure 1: Spike-encoding contig assembled from BioNTech/Pfizer BNT-162b2 vaccine. Although the full coding region is included, the nature of the methodology used for sequencing and assembly is such that the assembled contig could lack some sequence from the ends of the RNA. Within the assembled sequence, this hypothetical sequence shows a perfect match to the corresponding sequence from documents available online derived from manufacturer communications with the World Health Organization [as reported by https://berthub.eu/articles/posts/reverse-engineering-source-code-of-the-biontech-pfizer-vaccine/]. The 5’ end for the assembly matches the start site noted in these documents, while the read-based assembly lacks an interrupted polyA tail (A30(GCATATGACT)A70) that is expected to be present in the mRNA.
Auto-GPT
An experimental open-source attempt to make GPT-4 fully autonomous.
AutoRclone
A project that allows you copy large folders to Shared Drives.
Padavan-build
Padavan 自动编译
rclone_aria2_upload
wuweiluohua's Repositories
wuweiluohua/Auto-GPT
An experimental open-source attempt to make GPT-4 fully autonomous.
wuweiluohua/ccaa
Linux一键安装Aria2 + AriaNg + FileBrowse实现离线下载、文件管理。
wuweiluohua/ChatGPT-Telegram-Workers
Deploy your own Telegram ChatGPT bot on Cloudflare Workers with ease.
wuweiluohua/everyone-can-use-english
人人都能用英语
wuweiluohua/ExcelGPT
chatgpt for Excel ，基于ExcelDNA开发
wuweiluohua/ExplorerPatcher
This project aims to enhance the working environment on Windows
wuweiluohua/ExplorerPatcher-
ExplorerPatcher Chinese Localization - 在 Windows 11 上恢复高效的工作环境
wuweiluohua/gpt4free
decentralising the Ai Industry, just some language model api's...
wuweiluohua/GPT_API_free
Free ChatGPT API Key，免费ChatGPT API，支持GPT4 API（低价），ChatGPT国内可用免费转发API，直连无需代理。可以搭配ChatBox等软件/插件使用，极大降低接口使用成本。国内即可无限制畅快聊天。
wuweiluohua/healthcode-backup
备份一些健康码演示工具
wuweiluohua/How-To-Ask-Questions-The-Smart-Way
本文原文由知名 Hacker Eric S. Raymond 所撰寫，教你如何正確的提出技術問題並獲得你滿意的答案。
wuweiluohua/koreader
An ebook reader application supporting PDF, DjVu, EPUB, FB2 and many more formats, running on Cervantes, Kindle, Kobo, PocketBook and Android devices
wuweiluohua/leopard
On-device speech-to-text engine powered by deep learning
wuweiluohua/lookbusy-docker
wuweiluohua/magisk-files
wuweiluohua/Microsoft-Activation-Scripts
A Windows and Office activator using HWID / KMS38 / Online KMS activation methods, with a focus on open-source code and fewer antivirus detections.
wuweiluohua/Mr.-Ranedeer-AI-Tutor
A GPT-4 AI Tutor Prompt for customizable personalized learning experiences.
wuweiluohua/ms-ra-forwarder
Microsoft TTS for Legado Reader
wuweiluohua/NickelMenu
The easiest way to launch scripts, change settings, and run actions on Kobo eReaders.
wuweiluohua/SmartProxy
Firefox/Chrome browser extension. SmartProxy will automatically enable/disable proxy for the sites you visit, based on customizable patterns.
wuweiluohua/SwitchyOmega
Manage and switch between multiple proxies quickly & easily.
wuweiluohua/tachiyomi-extensions-
Source extensions for the Tachiyomi app.
wuweiluohua/tank
《蓝眼云盘》(Eyeblue Cloud Storage)
wuweiluohua/teledrive
The Google Drive/OneDrive/etc alternative using Telegram API
wuweiluohua/telegram-chatgpt-concierge-bot
Interact with OpenAI's ChatGPT via Telegram and Voice.
wuweiluohua/tgbot-book-
zhelper tgbot
wuweiluohua/TrackersListCollection
🎈 Updated daily! A list of popular BitTorrent Trackers. / 每天更新！全网热门 BT Tracker 列表！
wuweiluohua/v2rayN
wuweiluohua/Xiaomi-HyperOS-BootLoader-Bypass
A PoC that exploits a vulnerability to bypass the Xiaomi HyperOS community restrictions of BootLoader unlocked account bindings.
wuweiluohua/zlib-searcher
search zlib/libgen index to get ipfs_cid. We don't save and provide files, we provide search. I hope everyone have a copy of the index locally, so that no need to rely on any centralized service.