From 12163925b883ee7f844d52b37808c36773acd6c8 Mon Sep 17 00:00:00 2001 From: Alexander Lenail Date: Tue, 30 May 2023 05:59:36 -0400 Subject: [PATCH] add positive controls spreadsheet --- _pages/crispriacelllineprimer.md | 10 +++++----- compile_info.ipynb | 2 ++ resources/positivecontrolsgRNAlist_V4.xlsx | Bin 0 -> 14594 bytes 3 files changed, 7 insertions(+), 5 deletions(-) create mode 100644 resources/positivecontrolsgRNAlist_V4.xlsx diff --git a/_pages/crispriacelllineprimer.md b/_pages/crispriacelllineprimer.md index b01c21b..d1d8473 100644 --- a/_pages/crispriacelllineprimer.md +++ b/_pages/crispriacelllineprimer.md @@ -120,8 +120,8 @@ us to reserve GFP and mCherry for downstream experiments. To help labs test new CRISPRi cell lines, we have deposited on Addgene our published GFP CRISPRi reporter (Addgene: 46919). The GFP reporter allows you to generate single-cell flow cytometry data to troubleshoot and validate CRISPRi activity and cell line production. -We have included [positive control sgGFP sequences](http://weissmanlab.ucsf.edu/links/positivecontrolsgRNAlist_V4.xlsx) to be cloned into our sgRNA expression -vector (Addgene: 60955). We have also included with this protocol a list of [positive control sgRNAs](http://weissmanlab.ucsf.edu/links/positivecontrolsgRNAlist_V4.xlsx) +We have included [positive control sgGFP sequences](/resources/positivecontrolsgRNAlist_V4.xlsx) to be cloned into our sgRNA expression +vector (Addgene: 60955). We have also included with this protocol a list of [positive control sgRNAs](/resources/positivecontrolsgRNAlist_V4.xlsx) targeting human genes that have shown high activity across cell lines. We then use qPCR to measure knockdown efficiency for this second type of positive control. @@ -139,13 +139,13 @@ expression levels, we have found that we need to derive clonal cell lines and te CRISPRa activity using a functional assay.** To help labs test new CRISPRa cell lines, we have deposited on Addgene our published GFP -CRISPRa reporter (Addgene: Available soon). We have included [positive control sgRNA sequences](http://weissmanlab.ucsf.edu/links/positivecontrolsgRNAlist_V4.xlsx) +CRISPRa reporter (Addgene: Available soon). We have included [positive control sgRNA sequences](/resources/positivecontrolsgRNAlist_V4.xlsx) targeting the endogenous human CXCR4 gene. Both of these positive controls can be used to read out single cell CRISPRa activity using flow cytometry. We have also included -a [list of positive control sgRNAs](http://weissmanlab.ucsf.edu/links/positivecontrolsgRNAlist_V4.xlsx) targeting human genes that have shown high activity. We +a [list of positive control sgRNAs](/resources/positivecontrolsgRNAlist_V4.xlsx) targeting human genes that have shown high activity. We have used qPCR to measure gene activation efficiency for this second type of positive control. Additional protocols and reagents are linked here: -- [Positive control sgRNAs for CRISPRi and CRISPRa](http://weissmanlab.ucsf.edu/links/positivecontrolsgRNAlist_V4.xlsx) +- [Positive control sgRNAs for CRISPRi and CRISPRa](/resources/positivecontrolsgRNAlist_V4.xlsx) - [sgRNA cloning protocol](/resources/sgRNACloningProtocol.pdf) - [sgRNA assembly template](/resources/sgRNAAssemblyTemplate.xlsx) diff --git a/compile_info.ipynb b/compile_info.ipynb index 6a513b7..3fbb332 100644 --- a/compile_info.ipynb +++ b/compile_info.ipynb @@ -23,6 +23,7 @@ "cell_type": "markdown", "id": "a7486e04-b58e-49bd-97bd-e91e2862b221", "metadata": { + "jp-MarkdownHeadingCollapsed": true, "tags": [] }, "source": [ @@ -1232,6 +1233,7 @@ "cell_type": "markdown", "id": "ce98febf-fc16-4529-b8d6-2630646bb4fe", "metadata": { + "jp-MarkdownHeadingCollapsed": true, "tags": [] }, "source": [ diff --git a/resources/positivecontrolsgRNAlist_V4.xlsx b/resources/positivecontrolsgRNAlist_V4.xlsx new file mode 100644 index 0000000000000000000000000000000000000000..37342b5af958833449395bd0b4d6700a9f4a1c8f GIT binary patch literal 14594 zcmeHuV|ZmtyKS7T*y`AJ$F|XNI<{@w>e#mJj&0lQbl9<^&7`6bTLlgaQNw@>R&j+R@0`QAf$m z*2qDN&eh72AP)?LA_oZM0DN)Oepx*OPc}W zu?Y)h_UKHtps4Y`&L&JduYCdew8nWPS217owFN5~obYI>QR(KxADC$hRUaj31>m1w zJyN-(bW6qHLNH^pa^+9L$-JGP@GlW%h!7AAz*1 z4v3r@lR`3+1ru9U_FdwA6L*(vRRX?=Y*9^CWxI@TAA{7%F|);Ua|dJZf)26R%<;$y zBZ&3e6S1*)$|s7+iWq~Ru$;^J+F+DXxJUotXz%Z!K=S_*t@56-J=Y&kq(6cY<|A5l z?2RlP=;{7A|8JE34-U=0b-f~PN)nV2G33Jk0`YJ&(+V8PtO*0j9i6 zq1na;5f#JycAJqo-lp?q*qN{*Cc$vKC?DP?iHB$WXib5Gl4Sl9cUcP2@Z6CjJLA0IX(h_DDbB;R0 z!lrqsM>~WQdvM*OOH;#{?t|=jgh94kMfZU)q2ejKMGCoRJUeehq%sl&_gURz``FZl z@K7p^ks$FmRl=kMeLTr39Zr>TAz^@# z@^Du3_%R@2pg=&_Kv2N0mh}G-CN4Ji7Wy_e7Jt;V{|Xu4kHYqW@<03Nh?_9)V?YeN z2)@(|L+5h9I=D-01?B3iF$;qLxmjPR&vV-nTQ5u!LQ4KO=Z*$`Lgj z79-vqsrbPm0f)HSM1l10)(S~(Zv=ZRpk?*awUuc7R$egrY{By{E=@LRliTrZ7oeP- z@`N0AdeHN=A}Y4AV)etw6vJb%E>cX0FUG07tGHSkehqD)2pW~)7ph`f~E>?ELd z{!8CVs+LT(1!V{RxLTERaBn$~o3uivgVXTzMsp*dV3w2cfw1WS)6FnVTh@0UHXbvF zbu+IUApR9tw8=!2NLtzyk~_&{X#wNvC=&RlLtiU1%n_@elz0#^fmnTs7D3$J{tj@V zi~Eea!lFdOQ73|6z^-p6t*e;_i^LWPc^HREpKIZOAIZ=6-lNgyyT}k6nSS{ zG9IQtjOjSV{L0OCYTsVmkqGxbpb$OPE6cVc{|%Qqj{+J#l$CX&Sn^P(@(ivd6N;cp z-da@zY2{5NO@3DFV1e+D9*{m*(x+rp>2xN3~m0XVD3E4+k4{7ic#0R)HBZJK58Kl@(gMAXThygq|h>@HAz_ zT(-3agFA&Y2eL~K5nu2tmQs{}6+1q!nw13&(9;kmd%p)!2C{1CBIXmX5BNYcr?bNO zOs$OYMr>MsfATBW%w}Q{7dRjC#$an#On2XIr6%Bu&cCjYiNuG76dhE!p8mFBwWl5& zmvalz)l!A~sijGaXQ63q$v8nUWy*MiUyR?93Vk5kUxTk;Ci0F!a2JlT}BU_*e%!_e&~b*4?&Sna}tG5w(pw{%AgRR zHjV1933jY;$b6lt#esKLnI6oTXT7pM*IAz!Uzf?d0%U9A5kFd~9G1}Fk6&hd&bIsk)9$ zK0Bfh-*G4CE)V!ztPGLvLW4;2?526)U4fTVH;GCVE>>~+sae-^l$Gg^Mu|B=)n`OJ zWJ~V7)U6lbDPepzQit=fijwWxh)K^SLT(g}gNWtVBUaoP~!M=1bVeHSUJdxNVZ ze1*cM$aNZk0({N3@+n=Sh73>jXi`D{LaG`Pyc{&4RD~`DhIV5d*ij-GRVv2A+b}ib zLDc~RZM{KCg4$-KYJhYCb;XYirn08sl}&up?OEjU2*B>`mj#KrabqN-BjC zX5FK(yc{eys)PP?*KW!%=pN+*L;VTmQ*WR}_mE>;j4ih^d|=<(GLkZH_WP6`>1}t3 z1OQn@ilfl)kE_zUWch&k>O11go=#TxQ+H6G`Z@W96)*KFN4tixl>T$Ims_llLEf)pOVBMwt|hGXN1q4Zm$9Z)K;!=FRFgXL7V)T7G* z@sO+8dPHFa%c$SDgZvb?wyagRCvdL(gElHW^lk-rX`y@3{Y+S;^IBs$_H)TPZYG_n z!Z0aOTUrYu3g1-q!MhxCQNgT`4^ZjBhNQ(rg=UHZU%*SLoTcLCoB)>it+R!#k=*X- z?#cOPkA9h5kobaP+4Mh!5&bjT7+ z(5h8-Hs-7V`?khF0l!`g!)sSA-;K3pprodC*f=xSui821r!wAId-nYBPZmB>Mw~+Q zOceYkoq8WP0`$BO@MdUwc3sf@`AK;rer?xQM4XD|j{Qt0QSi)vr^oT3C;Lw9cgD z#5xXN0Vd|+$|Rw5qDkHvo$nid0sJO22$y)T8ftfLkloK6E;xEYB>ufQJRzd=%c#%aU!%5HYpgqVH#$gs37U=kI5phwdJP_2`kLzp(hVVw%avQ*G@IST zoulc<1P>G$$U5WKA7LbI;uO4>9xzY^4lPDe4>;pxG)*L~Gk(N%Qu46{ZKR;1j6+L=5*UhmoE%@*nl=az#TGw7r;SMO1B^D z-ySOzPzuAb?@PB|LhM{Z>{>$XUPA0yLhRjha`~J$c2IHKI9-={=JQG)^c4gWq~BY& z`_skD5o3qa)kGDXt}Dd`=4UwnkR@1Qb6MyPKB+Og{Ce+xr)Te!2@z1s_X#2Rhy+KvcV&S9M|-lWYBmi2A(`*1 z*DAc0#+*#B4G~L*3aoo;ok|n>n=u% z;MWgsoB~4VD$%|br<7pYmA>YYXio&fHRz?@5JeFw5S#l!m8hWAdR-DsJbA%#eVUy! 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